2,5,6-substituted N1 -alkylpyrimidines

ABSTRACT

The synthesis and application of N(1)-n-alkyl-pyrimidinium-salts are described. These surfactants have a very small critical micelle concentration (CMC) in the order of 10 -5  -10 -7  Mol/Liter. These N(1)-n-alkyl-pyrimidinium components have pharmacological activities and can act as antimetabolites.

The present invention relates to pharmaceutical preparations, knowncationic tensides as constituents of the pharmaceutical preparation, newchemical compounds (cationic tensides) which are used in particular asconstituent of the pharmaceutical preparations, processes for producingthe pharmaceutical preparation and processes for producing the known andnew chemical compounds (cationic tensides).

STATE OF THE ART AND ITS DISADVANTAGES

Micelles in aqueous solution, both non-ionic, cationic and anionic, havebeen described in the literature in numerous publications (Mittal, K. L.(1977) Micellization, Solubilization and microemulsions, Plenum Press,N.Y.--Mittal, K. L. (1979), Solution Chemistry of Surfactants, PlenumPress, N.Y.--Menger, F. M. (1977). In Bioorganic Chemistry III. Macro-and Multicomponent Systems (E. E. Van Tanelen, Ed.) Academic Press,N.Y.--Menger, F. M. (1979a) Acc. Chem. Res. 12, 111-117. On theStructures of Micelles.--J. H. Fendler, E. J. Fendler (1975) Catalysisin micellar and macromolecular Systems, Academic Press). Their structureand their galenical, medical and technical use is the subject ofnumerous investigations. Thus, the antiseptic effect of cetylpyridiniumchloride, benzethonium chloride and benzalkonium chloride or theirbromides is known. It is also known that in small concentrations theyexhibit bactericidal effect in vitro against a large number ofgrampositive and gramnegative bacteria, the gramnegative reactingsubstantially more sensitively than the grampositive. Also, certaingramnegative bacteria are resistant to these quaternary ammonium bases,e.g. Pseud. cepalia, Mycobact. tuberculosis.

Normally, cationic micelles in aqueous phase additionally have in theirhydrophobic core, which is largely defined by the aliphatic chain andits length, a hydrophobic-hydrophilic boundary layer (Stern layer) whichis hydrated and to some extent accommodates the counter ions. The sizeof this boundary layer is generally between 7-10 Å. They are alsosurrounded by the Guy-Chapman layer of 10-20 Å containingnon-electrostatically bound counter ions, e.g. Cl³¹ Br⁻, HSO₄ ⁻ andunstructured water. Only the concentrations of the counter ions andother ions effect a reduction of the critical micelle formationconcentration (cmc) at constant temperature, pressure and chemicalpotential, and the nature of the counter ions can govern the form andsize of the micelles in aqueous phase. This is done however only by thefraction of counter ions located in the Stern layer in the vicinity ofthe quaternary nitrogen.

The pure hitherto known cationic quaternary ammonium bases, officiallyalso referred to as invert soaps, have only a limited and non-specificantimicrobial effect (cf. e.g. W. Forth, D. Henschler, W. Rummel,Allgemeine und spezielle Pharmakologie und Toxikologie, 4th edition, B.I. Wissenschaftsverlag, 1983, p. 616). For this reason their use forexample as preservatives or disinfectants in the operative fields ofmedicine or in infection wards (antiseptics) is limited in spite oftheir low toxicity. Domagk recognised in 1935 cf. Wallhausser, K. H.:Sterilisation, Desinfektion, Konservierung, Keimidentifizierung,Betriebshygiene. 2nd edition, Thieme, Stuttgart 1978) that thequaternary ammonium bases are only bactericidally effective when atleast one of the substituents at the nitrogen consists of a linear alkylchain with 8-18 carbon atoms, the optimum chain length being C₁₂ -C₁₆.The best known representatives of this substance class are thebenzalkonium salts (chlorides and bromides). In addition,hexadecylpyridinium chloride and benzethonium chloride are known andhave achieved medical and pharmaceutical significance. The effect ofthese invert soaps depends of course very greatly on their environment.By soaps for example the effect is largely cancelled as it is also inthe acidic pH range. Blood, pus, stools and dirt likewise lead toinactivation. Moreover they have a protein-precipitating action whichstarts even at low concentrations of the N⁺ tensides, i.e. in the rangeof 1-2% by weight of aqueous solutions. At a concentration of theseknown tensides amounting to only 2-3 times the critical cmc, although noprotein precipitating effect (denaturing) occurs, a reversibleinactivation does take place of enzyme systems and support proteins byunfolding of the active three-dimensional structure ("loss of activitythrough unfolding").

Also known are the antibacterial and non-specific effect of quaternaryammonium compounds and their surfactant effect, of dequalinium acetate,cetyldimethylammonium bromide (CTAB) and hexadecylpyridinium chloride(CPCl), (cf. e.g. Goodman and Gilman's, The Pharmacological Basis ofTherapeutics, EDS. A. G. Goodman, L. S. Goodman, Th. W. Rall, F. Murad,1985, 7th Edition, Collier, MacMillan Publishing Company, N.Y., p. 971;Merck Index, 1985). The micellar properties of these compounds have beenrelated to their surface activity and antimicrobial properties (cf.Attwood, D, and Florence, A. T., Surfactant Systems, Chapman and Hall,London and N.Y., 1983). However, the non specific surface activity ofthese quaternary aliphatic and aromatic ammonium bases cannot beregarded a priori as prerequisite for the antibacterial, antifungal andkeratolytic effect because nonionic detergents, e.g. Brij, Triton X 100,Lubrol etc. do not become reactive.

Organic quaternary ammonium bases of the type (R_(n), R₁, R₂, R_(m),N⁺)Y⁻ (HET=N⁺ --(CH₂)_(x) --CH₃)Y⁻ and [(H₃ C)₃.C C--CH₂ --C(CH₃)₂ --X₁--[O--(CH₂)₂)₂ --N⁺ (CH₃)₂ --CH₂ --X₂ ]Y⁻ are only partially known, e.g.hexadecyltrimethylammonium chloride and bromide(cetyltrimethylammonium), hexadecylpyridinium chloride or bromide(cetylpyridinium chloride) and N,N'-dimethyl-N-22-4-(1,1,3,3-tetrymethylbutyl)phenoxyethylphenylmethanium chloride(benzethonium chloride, methylbenzethonium chloride) and thebenzalkonium chlorides with alkyl radicals of C₈ H₁₇ to C₁₈ H₃₇. Theseknown N⁺ tensides all have a small critical micelle formation constant(cmc) in the range of 10⁻⁴ -10⁻⁵ mol, depending on the environmentalconditions such as ionic strength, temperature, pressure and addition oforganic solvents of specific dielectric constants. The influence of ananion, Y⁻, and of fractionated bonds, number of anions at the micellesurface (Stern layer) and their influence on the geometric form of theoverall cationic micelle of the aforementioned quaternary organicammonium bases, have so far been the subject of little investigation.This also applies to the form of the aforementioned tensides in thepresence of potentiating mixtures, such as glycerol, dimethyl sulfoxideethanol, propanol and their stability to temperature and absorptivecapacity for hydrophobic (lipophilic) pharmaceutical active substances.Here, no quantifiable investigations are available for theaforementioned N⁺ tensides either.

Tensides of the general formula (HET=N⁺ --(CH₂)_(x) --CH₃)Y⁻, theheterocycle being a benzimidazole, pyrimidine, imidazole, thiazole,benzthiazole or purine radical have so far not been described, and norhas their micelar behaviour in aqueous solutions in the presence andabsence of potentiating mixtures. This applies equally to substitutedpyridinium compounds which in addition, as will be shown later, can formin aqueous solution vesicles of specific size and form.

The relatively broad and undifferentiated effect mechanism of thealready known quaternary organic ammonium bases and the resulting fieldof use as antiseptics and their toxic action at higher therapeuticaldoses has restricted the pharmaceutical use of these organic quaternaryammonium bases. Also, for 1% by weight or higher concentrations inaqueous solutions, creams and ointments hypersensitive, allergic andtopical irritations have been observed so that specific therapeuticaluse is possible only to a limited extent.

The bactericidal effect of chlorhexidine is known in the case ofgrampositive and gramnegative bacteria but gramnegative bacilli areresistant.

Pharmaceutic preparations permitting a more specific therapy withpharmaceutical active substances included in micelles, e.g. ofantiviral, antifungal, antineoplastic nature, are not available intherapeutically effective doses and a suitable pharmaceuticalpreparation (galenic).

A great disadvantage of the hitherto known pharmaceutical preparationsof quaternary organic ammonium bases, this applying in the presence ofpotentiating mixtures as well, is the polydispersity of the colloidalmicellar solutions. Depending on the pharmaceutic preparation form, pHvalue, ionic strength, counter ion Y⁻ and temperature, hitherto in apharmaceutical preparation micelles of various form and size andstability and absorptive capacity for pharmaceutical active substanceswere present.

In the broadest sense micelles are taken to mean aggregates of dissolvedmolecules formed by association. In the narrower sense mainly used todaymicelles is a term applied to aggregates which form from tensidemolecules -n aqueous solutions above a specific temperature Kraft point)or a characteristic concentration. This concentration is called thecritical micellization concentration, cmc. When the cmc is exceeded themonomer concentration remains practically constant and the excesstenside molecules form micelles. They may occur in various shapes(spheres, rods, discs) depending on the chemical constitution of thetenside and on the temperature, concentration or ionic strength of thesolution. The micelles have characteristic aggregation numbers withusually only a small distribution spread. Reaching the cmc manifestsitself by abrupt changes in the surface tension (which is utilized tomeasure the cmc), the osmotic pressure, the electrical conductivity andthe viscosity.

Micelles are thermodynamic stable association colloids of surfactantsubstances in which the hydrophobic radicals of the monomers lie in theinterior of the aggregates and are held together by hydrophobicinteraction (van-der-Waals forces); the hydrophilic groups face thewater and by solvation provide the solubility of the colloid.

Further information on micelles will be found in Rompps Chemielexikon,8th edition, Franckh'sche Verlagsbuchhandlung Stuttgart, 1985, page 2600et seq.

An object of the present invention is to provide a pharmaceuticalpreparation which contains the active substance in the most stable formpossible and in which the active substance is liberated at the locationof the pathological process as rapidly and completely as possible.

This problem is solved according to the invention by a pharmaceuticalpreparation which is characterized in that it is made up of a micelleconsisting of a cationic tenside with a monovalent anion and ahydrophobic pharmaceutical active substance dispersed in a solvent whosepH value is ≦7, the critical micellitation concentration (cmc) lying inthe range of 1.0.10⁻⁷ to 1.5.10⁻⁴ mol/liter.

Preferably, this pharmaceutical preparation is made up of a micelleconsisting of a cationic tenside with a monovalent anion in an amount of0.01 to 0.1% by weight with respect to the total pharmaceuticalpreparation, and a hydrophobic pharmaceutical active substance in anamount of 0.001 to 0.5% by weight with respect to the totalpharmaceutical preparation, dispersed in a solvent whose pH value is≦7.0 , in an amount of 99.40 to 99.989% by weight with respect to thetotal pharmaceutical preparation, the critical micellizationconcentration (cmc) lying in the range of 1.0.10⁻⁷ to 1.5.10⁻⁴mol/liter.

The micelles described here in aqueous phase have with a hydrophobicchair length of 15--(CH₂) groups including their quaternary nitrogen inthe aromatic structure a diameter of approx. 50-100 Å depending on thenature of the counter ions.

DESCRIPTION AND PREPARATION OF THE QUATERNARY AMMONIUM BASES

The cationic tenside according to the invention is preferably a compoundof the general formula ##STR1## wherein preferably

R₁ =an alkyl radical with 1-12 C atoms or an aralkyl

R₂ =an alkyl radical with 1-12 C atoms or an aralkyl

R_(n) =a straight-chain or branched alkyl radical, which may besubstituted, with 1-22, preferably 10-20 C atoms or an alkenyl radicalwith 8-20 C atoms, preferably 8-10 C atoms or a 5- or 6-member aromaticheterocycle with one or 2 nitrogen atoms and optionally one sulfur atomor one oxygen atom and

R_(m) =a straight-chain or branched alkyl radical, which may besubstituted, with 1-22, preferably 10-20 C atoms or an alkenyl radicalwith 8-20 C atoms, preferably 8-10 C atoms or a 5- or 6-member aromaticheterocycle with one or 2 nitrogen atoms and optionally one sulfur atomor one oxygen atom, or a quinolinium radical, and

Y⁻ =a monovalent anion.

Further preferred embodiments are:

The straight-chain or branched alkyl are preferred to be those with C₆-C₂₂, in particular however C₁₂ -C₂₀, carbon atoms, for examplen-heptyl, 2-methylhexyl, 3-methylhexyl, 3-ethylpentyl, 2,2, 2,3, 2,4, or3,3-dimethylpentyl, n-octyl, 4-methylheptyl, 2,2,2, 2,2,4, 2,3,3,2,3,4-trimethylpentyl, n-nonyl, n-decyl, n-undecyl, n-dodecyl,n-tridecyl, n-tetradecyl, n -pentadecyl, n-hexadecyl (cetyl),n-heptadecyl, n-octadecyl, n-nonadecyl, or n-eicosyl (arachinyl).

Preferred is a straight-chain alkyl having an even number of 10-20carbon atoms, e.g. n-dodecyl, n-tetradecyl, n-hexadecyl (cetyl),n-octadecyl or n-eicosyl. They all have the same bonding and absorptivecapacity for inorganic and organic (hydrophobic) active substances, forexample Hg(CN)₂, ZnEDTA, ZnO, and K₁₈ (KW₂₁ Sb₉ O₈₆)₁₇ as inorganicantiviral active substances, and azathioprine, nystatin, amphotericin,idoxuridine, cytarabine and trifluorothymidine as organic activesubstances.

Preferred is an alkenyl having 12-20 carbon atoms for R_(n) if R_(m) isa methyl, ethyl up to a hexyl radical, specifically alkenyl, having adouble bond, such as 9-cis-dodecenyl, 9-cis-tetradecenyl,9-cis-hexadecenyl, 6-cis-octadecenyl, 6-trans-octadecenyl and9-cis-octadecenyl.

R₁, R₂ and R_(m) is preferred to be methyl, ethyl or also hexyl.

An aromatic heterocycle for R_(n) of the formula (I) is a 5 or 6-memberaromatic heterocycle having one or two nitrogen atoms, and optionally anitrogen and a sulfur atom, e.g. a pyridine, a pyrimidine, a pyrazine(1,4-diazine), a pyrazole, an imidazole, a thiazole and purine radical(7N-imidazolium [4,5-d]pyrimidine) or a benzo-condensed thiazole andimidazole radical, e.g. N₃ -benzimidazole or benzthiazole.

Substituents of this heterocycle are at the nitrogen atom and possiblyat a carbon atom at a low alkyl, e.g. methyl or ethyl, or a hydroxy lowalkyl, e.g. hydroxymethyl or 2-hydroxyethyl, oxo, hydroxy or halogen,e.g. chlorine or bromine.

A heterocycle is preferably 2 or 4-low alkyl pyridinium, e.g. 2 or4-methyl or 2 or 4-ethylpyridinium, di-low alkyl pyridinium, e.g.2,6-dimethyl, 2-methyl-3-ethyl, 2-methyl-4-ethyl, 2-methyl-5-ethyl or2-methyl-6-ethylpryridinium, 2, 3 or 4-halogen pyridinium, e.g. 2, 3 or4-chloropyridinium or 2, 3 or 4-bromopyridinium, 2-low alkylimidazolinium, oxazolinium or thiazolinium, e.g. 2-methyl or 2-ethylimidazolinium, oxazolinium or thiazolinium or 2-low alkyl-8-halogenquinolinium, e.g. 2-methyl-8-chloroquinolinium.

Y.sup.⊖ is an anion, preferably chloride, bromide, iodide or ethylsulfate, a low alkonate, such as formate acetate, propionate, hydrogensulfate (HSO₄ --), malate or fumarate, salicylate, alginate orgluconate.

A cationic tenside of the general formula (I) is preferablyN-benzyl-N,N-dimethyl-N-2-[2-(4-(1,1,3,3-tetramethylbutyl)-phenoxy)-ethoxy]-ethylammoniumchloride,N-benzyl-N,N-dimethyl-N-2[2-(3-methyl-4-(1,1,3,3-tetramethylbutyl)-phenoxy)-ethoxy]-ethylammoniumchloride (methylbenzethonium chloride), n-dodecyltrimethylammoniumchloride or bromide, trimethyl-n-tetradecylammonium chloride or bromide,n-hexadecyltrimethylammonium chloride or bromide (cetyltrimethylammoniumchloride or bromide), trimethyl-n-octadecylammonium chloride or bromide,ethyl-n-dodecyldimethylammonium chloride or bromide,ethyldimethyl-n-tetradecylammonium chloride or bromide,ethyl-n-hexadecyldimethylammonium chloride or bromide,ethyldimethyl-n-octadecylammonium chloride or bromide,n-alkyl-benzyldimethylammonium chloride or bromide (benzalkoniumchloride or bromide), e.g. benzyl-n-dodecyldimethylammonium chloride orbromide, benzyldimethyl-n-tetradecylammonium chloride or bromide,benzyl-n-hexadecyldimethylammonium chloride or bromide orbenzyldimethyl-n-octadecylammonium chloride or bromide,N-(n-decyl)-pyridinium chloride or bromide, N (n-dodecyl)-pyridiniumchloride or bromide, N-(n-tetradeyl)-pyridinium chloride or bromide,N-(n-hexadecyl)-pyridinium chloride or bromide (cetylpyridiniumchloride) or N(n-octadecyl)-pyridinium chloride or bromide or a mixtureof these tensides.

A cationic tenside of the general formula (I) R_(n) N.sup.⊕ (R₁,R₂)R_(m)Y.sup.⊖ is preferably with R_(n) =R₁ =R₂ e.g. R_(n) N.sup.⊕ (CH₃)₃Y.sup.⊖ or as substance e.g. n-heptyl-trimethyl-ammonium chloride(bromide), 3-methyl-hexyl-trimethyl-ammonium chloride,n-nonyl-trimethyl-ammonium chloride, n-undecyl-trimethyl-ammoniumchloride, n-hexadecyl-trimethyl-ammonium chloride, n-octadecyl orn-eicosyl-trimethyl-ammonium bromide with an even number of 12-20 carbonatoms.

On the basis of a microemulsion and/or ointment e.g. in the presence ofup to 10% (^(g) /g) DMSO these N tensides have the same antifungal,antibacterial and keratolytic properties as the non-covalently boundpharmaceutical active substances.

The tensides of the general formula R_(n) N.sup.⊕ (R₁,R₂)R_(m) Y.sup.⊖are to be prepared analogously to that described in the standard work"Cationic Surfactants" by E. Jungermann, Dekker, N.Y., 1970, cf. alsothe handbook which appears each year "McCutcheon's Emulsifiers andDetergents" Manufacturing Confectioner Publishing Co. Other alkylpyridinium halides can be obtained by reaction of stoichiometric amountsof pyridine derivatives with long-chain alkyl halides in good yield.Other processes proceed from the corresponding ultracyclic N-compoundsand 1,3-propane methane, as for example described in F. J. Fendler etal., J.Chem.Soc., Perkin III, 1097 (1977). Other processes leading tosimilarly good yields are for example described in Attwood, D.,Elwarthy, P. H., and Kaye, S. B., J.Phys.Chem. 74, 3529 (1970) and maybe used analogously for the synthesis of the substances of formula II.The pharmaceutical active substances are available commercially.

The synthesis of the compounds of the general formula R_(n), R_(m), R₁,R₂ N.sup.⊕ Y.sup.⊕ or R_(n), R_(m) N.sup.⊖ (CH₃)₂ Y.sup.⊖ is carried outspecifically in accordance with the following procedure:

(a) The corresponding alkyl iodide or bromide is allowed to stand withan excess of trimethylamine (halide:amine=1:1.45) for 24 hours at 20° C.in an autoclave for preparing the corresponding quaternary ammoniumbase. No solvent other than methanol which has been saturated with thetrimethylamine or R₁, R₂ alkylamine was used. The reaction mixture isstirred into 5 times the volume of ether and heated in reflux for 20min. The solid residue forming after cooling in ether is filtered off.The recrystallization is from chloroform. The crystals are washedrepeatedly with anhydrous ether. The recrystallizations until constantmelting point were carried out from ethanol/ether (1:1, % ^(g) /g) inthe presence of activated charcoal. The crystals were dried overnight at80° C. over calcium chloride under vacuum at 1 mm/Hg.

(b) To prepare R_(n), R_(m).sbsb.⊕, R₁,R₂ N.sup.⊕ Y.sup.⊖ thecorresponding amines, R₁, R₂ --N.sup.⊕ -amines, were refluxed with thestoichiometric amounts of R_(n), R_(m) -iodides in absoluteethanol-hexane (1:2 % ^(g) /g) for 48 hours. Thereafter the reaction wascooled and the mixture poured into a 5-times excess of ether andfiltered off. The recrystallization was carried out as indicated under(a).

(c) To convert the quaternary ammonium halides to the correspondingbromides, chlorides or also iodides, the following methods are possible:300 g Amberlite IRA-400 (4 mequiv/g) in the chloride form is introducedinto a column (45×5 cm) and with a very slow throughflow time washedwith 1 liter of a 20% aqueous solution of potassium chloride orpotassium bromide or potassium iodide or KY.sup.⊖. The matrix was thenwashed with deionized water until no reaction occurred to chloride orbromide or iodide.

Thereafter the column matrix was charged with a 10% aqueous solution ofa quaternary ammonium bromide. The following elution was carried outwith water with a flow rate of 1 ml/min. The corresponding quaternaryammonium bromide or halide was obtained by concentration of the eluatein a rotary evaporator. The recrystallization was carried out asdescribed under (a). The following table 4 shows some cationic tensidesof the form R_(n) N.sup.⊕ (CH₃)₃ Y.sup.⊖ which have been prepared bythis process.

A subclass of the compounds of the general formula (I) is the compoundof the general formula ##STR2## These are derivatives of thebenzethonium halides. By substitution of the radicals X₁ and X₂, whereX₁ may be equal to X₂, these compounds can be made analogously asalready described in U.S. Pat. No. 2,115,250 (1938) or U.S. Pat. No.2,170,111 (1939) and 2,229,024 (1941). These specific N-tensides areparticularly stable even in the presence of a potentiating mixture andsurprisingly have a high absorptive capacity for micellar inclusion ofpharmaceutical active substances. Moreover; when carried out accordingto this method they are independent of the environment. Y.sup.⊖ is ananion, for example chloride, bromide or also iodide, a low alkonate,such as formate, acetate, propionate, malate or fumarate, salicylate,alginate or gluconate.

                                      TABLE 4                                     __________________________________________________________________________    Preparation and melting point and elementary analysis                         of the quarternary ammonium compounds of the type                             RN.sup.+ (CH.sub.3).sub.3 Y.sup.⊖                                     from R.sub.n, R.sub.m, R.sub.1, R.sub.2 N.sup.⊕ Y.sup.⊖           with R.sub.1 ═R.sub.2 and R.sub.n ═R.sub.m.                                         cmc        Analysis found                                       No. R       Y.sup.⊖                                                                 mol   Fp. °C.                                                                     C  H  N  Y                                           __________________________________________________________________________     1  Methyl  Br                                                                              1,5 × 10.sup.-5                                            2  Ethyl   I 2,0 × 10.sup.-5                                                               >300.sup.c                                                                         27,90                                                                            6,56                                                                             6,49                                                               >300.sup.d                                                                         27,92                                                                            6,56                                                                             6,51                                            3  n-Propyl                                                                              I 2,0 × 10.sup.-5                                                               190  31,51                                                                            7,05                                                                             6,09                                                               189  31,46                                                                            7,04                                                                             6,11                                            4  Isopropyl                                                                             I 3,5 × 10.sup.-5                                                               >300 31,50                                                                            7,08                                                                             6,09                                                               316  31,46                                                                            7,04                                                                             6,11                                            5  n-Butyl I 4,1 × 10.sup.-5                                                               231  34,69                                                                            7,48                                                                             5,72                                                               226  34,58                                                                            7,46                                                                             5,76                                            6  t-Butyl I 6,0 × 10.sup.-6                                                               256  34,66                                                                            7,47                                                                             5,72                                                               260  34,58                                                                            7,46                                                                             5,76                                            7  n-Pentyl                                                                              I 7,0 × 10.sup.-5                                                               224  37,28                                                                            7,86                                                                             5,41                                                                    37,37                                                                            7,84                                                                             5,45                                            8  l-Methylbutyl                                                                         I 1,0 × 10.sup.-6                                                               224  37,48                                                                            7,87                                                                             5,43                                                                             49,17                                                                37,37                                                                            7,84                                                                             5,45                                                                             49,34                                        9  n-Hexyl I 7,9 × 10.sup.-6                                                               160  39,68                                                                            8,19                                                                             5,11                                                               166  39,86                                                                            8,18                                                                             5,16                                           10  Cyclopentyl                                                                           I 6,0 × 10.sup.-6                                                               271  37,78                                                                            7,13                                                                             5,41                                                                             49,63                                                                37,66                                                                            7,11                                                                             5,47                                                                             49,74                                       11  Cyclohexyl                                                                            I 7,1 × 10.sup.-6                                                               271  40,25                                                                            7,48                                                                             5,18                                                                    40,16                                                                            7,49                                                                             5,20                                           12  Allyl   I 1,5 × 10.sup.-7                                                               104  31,81                                                                            6,22                                                                             6,15                                                                             55,76                                                           102  31,73                                                                            6,21                                                                             6,17                                                                             55,89                                       13  2-Propynyl                                                                            I 6,0 × 10.sup.-5                                                               181  32,09                                                                            5,40                                                                             6,19                                                                             56,29                                                                32,01                                                                            5,37                                                                             6,22                                                                             56,39                                       14  3-Butenyl                                                                             I 3,5 × 10.sup.-5                                                               236  34,93                                                                            6,70                                                                             5,78                                                                             52,56                                                                34,87                                                                            6,69                                                                             5,81                                                                             52,63                                       15  Phenyl  I 7,0 × 10.sup.-5                                                               227  41,12                                                                            5,38                                                                             5,31                                                                             48,15                                                           227  41,08                                                                            5,36                                                                             5,32                                                                             48,23                                       16  Benzyl  I 7,3 × 10.sup.-5                                                               179  43,33                                                                            5,82                                                                             5,00                                                               179  43,33                                                                            5,82                                                                             5,05                                           17  4-Chlorbutyl                                                                          I 5,1 × 10.sup.-6                                                               182  29,42                                                                            5,97                                                                             5,01                                                                    30,28                                                                            6,17                                                                             5,05                                           18  4-Brombutyl                                                                           I 7,0 × 10.sup.-6                                                               131  25,30                                                                            5,40                                                                             4,62                                                                    26,10                                                                            5,32                                                                             4,35                                           19  4- Iodobutyl                                                                          I 1,5 × 10.sup.-7                                                               160  23,43                                                                            4,75                                                                             4,00                                                                             67,80                                                                22,78                                                                            4,64                                                                             3,79                                                                             68,79                                       20  2-Ethoxyethyl                                                                         Br                                                                              2,0 × 10.sup.-7                                                               174  39,07                                                                            8,44                                                                             6,49                                                                             38,48                                                                39,63                                                                            8,55                                                                             6,60                                                                             37,67                                       21  2-Phenoxyethyl                                                                        Br                                                                              1,5 × 10.sup.-7                                                               162  50,74                                                                            6,98                                                                             5,34                                                                             30,79                                                                50,78                                                                            6,97                                                                             5,38                                                                             30,71                                       22  p-Methylbenzyl                                                                        Br                                                                              2,0 × 10.sup.-7                                                               197  53,97                                                                            7,78                                                                             5,66                                                                             32,49                                                                54,10                                                                            7,43                                                                             5,74                                                                             32,72                                       23  p-Fluorbenzyl                                                                         Br                                                                              2,5 × 10.sup.-7                                                               237  48,32                                                                            6,10                                                                             5,61                                                                    48,40                                                                            6,09                                                                             5,65                                           24  p-Chlorbenzyl                                                                         Br                                                                              3,0 × 10.sup.-5                                                               207  45,39                                                                            5,71                                                                             5,29                                                                    45,39                                                                            5,75                                                                             5,26                                           25  p-Brombenzyl                                                                          Br                                                                              4,0 × 10.sup.-5                                                               220  38,93                                                                            4,92                                                                             4,52                                                                             51,59                                                                38,86                                                                            4,89                                                                             4,53                                                                             51,71                                       __________________________________________________________________________

The cationic tenside according to the invention is preferably a compoundof the general formula

    [HET=N.sup.+ --(CH.sub.2).sub.x --CH.sub.3 ]Y.sup.-

wherein

HET=N⁺

is a substituted or non-substituted pyridinium radical or

a substituted or non-substituted pyrimidinium radical or

a substituted pyrazine-(1,4-diazinium) radical or

an imidazolium radical (4,5-d)pyrimidine radical, substituted ornon-substituted, or

a substituted or non-substituted imidazolium radical or

a substituted or non-substituted pyrazolium radical or

a substituted or non-substituted thiazolium radical, or

a substituted or non-substituted benzthiazolium radical or

a substituted or non-substituted benzimidazolium radical,

x=8 to 20 and

y⁻ =chloride, bromide, iodide, formate, acetate, propionate, hydrogensulfate, malate, fumarate, salicylate, alginate, gluconate or ethylsulfate.

Preferred embodiments of this cationic tenside are the followingcompounds:

In the following embodiments, in which y⁻ occurs, this y⁻ denotes ineach case one of the above thirteen anions.

N-alkyl pyridinium of the formula ##STR3## hexadecylpyridinium of theformula ##STR4## N-alkyl-4-hydroxypyridinium of the formula ##STR5##hexadecyl-4-hydroxypyridinium of the formula ##STR6## 2,5,6 substitutedN₁ -alkyl pyridinium compounds of the formula ##STR7## 2,5,6 substitutedN₁ -hexadecylpyrimidinium of the formula ##STR8##4-n-alkyl-pyrazinium-2-carboxamide of the formula ##STR9##4-hexadecylpyrazinium-2-carboxamide of the formula ##STR10##7-n-alkyl-imidazolium [4,5-d]-pyrimidine of the formula ##STR11##7-hexadecylimidazolium [4,5-d]pyrimidine of the formula ##STR12##3-n-alkyl-5,6-substituted benzimidazolium compounds of the formula##STR13## 4-substituted 2-hexadecylpyrazinium compounds of the formula##STR14## 1-n-alkyl-4-substituted imidazolium compounds ##STR15##1-hexadecyl-4-substituted imidazolium compounds of the formula ##STR16##3-n-alkyl-5,6-substituted thiazolium compounds of the formula ##STR17##3-n-hexadecyl-2,5-substituted thiazolium compounds of the formula##STR18## 3-n-alkyl-5,6-substituted benzthiazolium compounds of theformula ##STR19## 4-[1,1 bis n-alkyl (low alkyl)]N-hexadecylpyridiniumcompounds of the formula ##STR20## 3,5 bis[(n-alkyloxy)carbonyl]N-hexadecylpyridinium compounds of the formula##STR21## 4-(17-tritriacontyl)-n-methyl-pyridinium chloride of theformula ##STR22## 3,5 bis [(n-hexadecyloxy)carbonyl)]-N-methylpyridinium chloride ##STR23## Cationic tensides ofthe general formula ##STR24## wherein

x₁ =a non-substituted phenyl radical or a phenyl radical substituted inthe 4-position or in the 3.5-position or in the 1,2,4,5-position,

x₂ =a non-substituted phenyl radical or a phenyl radical substituted inthe 4-position or in the 3,5-position or in the 1,2,4,5-position and

y⁻ =a monovalent anion.

GENERAL REMARKS ON THE THE PREPARATION OF THE (HET=N⁺ -(CH₂)_(x) --CH₃)Y⁻ compounds II:

The cationic tensides according to the invention of the general formulaII are novel apart from hexadecylpyridinium halide.

In the cationic tenside of the general formula II HET=N⁺ is preferably asubstituted or non-substituted pyridinium radical or a substituted ornon-substituted pyrimidinium radical or a substitutedpyrazine-(1,4-diazinium) radical or an imidazolium radical (4,5-d)pyrimidine radical, substituted or non substituted, or a substituted ornon-substituted imidazolium radical or a substituted or non-substitutedpyrazolium radical, or a substituted or non-substituted thiazoliumradical or a substituted or non-substituted benzthiazolium radical, or asubstituted or non-substituted benzimidazolium radical.

These cationic tensides are characterized in that they have a very smallcritical micellization constant (cmc) of approximately 1.5×10⁻⁷ mol, arevery highly antimicrobial and antifungal, do not exhibit anypolydispersity in the presence of inorganic anions or potentiatingmixtures and in some cases themselves are microbial metabolism products(antimetabolites) which are not toxic for the host cell.

The formation of the salt-like structure of this class of cationictensides of the form (HET=N.sup.⊕ --(CH₂)_(x) --CH₃) Y.sup.⊕ is interalia due to the electron density distribution of the heteroaromaticcores and their basicity, including the influence of the substituents. Anecessary condition leading to the formation of quaternary salts of thisfive and six-member heteroaromatic class is that the electron density atthe nitrogen which is rendered quaternary has a magnitude determined byMO-SEF calculations of -0.03 (e.g. pyrazine-N₄) to -0.159 (e.g.imidazole-N₁, purine N₇) The stability of the individual heterocycliccationic tensides described here is moreover also governed by theirsymmetry and the length of the alkyl chain at the quaternary nitrogen:

In the case of the imidazole, benzimidazo-e, for example, stabilizationis by formation of the salt at the quaternary nitrogen N₁ and the freeelectron pair at N₃ and the resulting high symmetry. The same applies tothe H₉ -tautomer of purine and its symmetrically arranged substituentswhich influence the negative charges at the N₁ (-0.124), N₃ (-0.108) andN₉ (-0.149) such a manner that the quaternization at the N₉ is preferredin that the aforementioned order N₁ →N₃ →N₉ is reversed. The yields canbe increased by the choice of suitable solvents. Whereas for pyridine,pyrimidine and imidazole radicals symmetrical effects at the core playan important part in the case for example of pyrazine the electroniceffect in the 2-position is significant but there are also verypronounced inductive effects (e.g. 2-amino group), less than mesomers.This also applies to pyrazole.

The length of the alkyl chain at the quaternary nitrogen atom governsnot only the melting point and hydrophobicity of the cationic micellessubsequently formed in aqueous solutions; in addition, the yieldsdecrease with increasing chain length whilst the reaction times increasefor example in nitrobenzene of 2-ethoxyethanol. Stable and easilycrystallizable compounds are obtained for C₁₂ -C₁₈, the counter ionY.sup.⊖ being without exception bromide and chloride. The othercompounds can easily be recrystallized from acetone or chloroform. Thecorresponding iodine compounds are sensitive to heat and light.

SPECIFIC PREPARATION OF THE (HET=N.sup.⊕ --(CH₂)_(x) --CH₃) Y.sup.⊕compounds

(a) The corresponding compounds of pyridine or substituted Pyridine, assix-member heterocycle, can be prepared from the corresponding alkylbromides or iodides in methanol at 35° C. and pyridine or substitutedpyridines with a yield of 70%. The corresponding molar amounts of thealkyl bromide, almost all of which are available commercially but whichmust be subsequently preparatively purified by high-pressure liquidchromatography (HPLC), are firstly dissolved in methanol (1( timesexcess volume with respect to pyridine) and under nitrogen thestoichiometric amount of pyridine, also dissolved in methanol, addeddropwise whilst stirring. Heating is carried out for 6 hours underreflux whilst stirring at 70° C. so that the reaction yield is almostquantitative. Thus, for example, the yield ofhexadecyl-4-hydroxypyridinium chloride or bromide in methanol as solventis 95%, with ethanol 80% and in ether/ethanol only 40%.

Dodecylpyridinium chloride is obtained with a yield of almost 70%. 3,5dihydroxydodecylpyridinium bromide is formed quantitatively inaccordance with the above procedure from dodecyl bromide and3,5-dihydroxypyridine in boiling chloroform after 4 hours (melting point180° C.).

Purification of the corresponding pyridinium compounds: By repeatedrecrystallization from mixtures of methanol/ether, starting with ⁴⁰/60(^(v) /v); ⁵⁰ /50(^(v) /v) and finally ⁹⁰ /10(^(v) /v), the desiredproducts are obtained with constant melting point, uniform molecularweight and specific surface-active properties (measured by theconcentration dependence of the surface tension). In addition thesecompounds exhibit the typical ¹ H-NMR signals outlined above. Thenumerous CH₂ groups and the CH₃ group generate a clearly visibleabsorption band in the IR spectrum at 2930 cm⁻¹ and 2850 cm⁻¹ (methylenegroup) a medium-weak band at 2960 cm⁻¹ and a weak band at 2870 cm⁻¹which can be assigned to the methyl group. A rapid and quantitativeseparation cf the n-alkyl pyridinium halides from unconverted n-alkylbromides and pyridine is achieved by preparative high-pressure liquidchromatography on an RP18 column with the aid of an elution mixtureconsisting of 60% (^(v) /v) methanol (ethanol) and acetone nitrile 40%(^(v) /v) isocratic at 9.52 atm column pressure (Uv detection at 260nm).

(b) Pyrimidine compounds

(1) Hexadecylpyrimidinium bromide, 0.01 mol, 5-aminopyrimidine (0.95 g)and hexadecyl bromide, 0.01 mol (3.05 g), are reacted in 20 ml methanolwhilst stirring under nitrogen at 20° C. for 24 hours in the presence ofcatalytic amounts (0.5 mg) sodium amide. The resulting N₁-hexadecyl-5-aminopyrimidinium bromide i: dissolved in acetone at 76° C.and after cooling to room temperature the N₁ -hexadecyl5-aminopyridinium bromide crystallizes with a melting point of 122° C.Yield 35%.

0.01 mol of this N₁ -hexadecyl-5-aminopyrimidinium bromide (3.20 g) arestirred in methanol/water ⁵⁰ /50 (^(v) /v) at 0° C. in an ice bath with1 g NaNO₂ and 0.1 ml concentrated hydrobromic acid under nitrogen for 6hours. Thereafter the mixture is brought to room temperature andsubsequently refluxed at 80° C. for 2 hours under nitrogen whilststirring. The resulting hexadecylpyrimidinium bromide is extracted with2-ethoxyethanol and caused to crystallize out at 10° C. Yield 30%,melting point 105° C. (bromide), 189° C. (chloride).

Preparative separation of non-converted products can also be achieved byhigh pressure liquid chromatography as described for the pyridiniumderivatives.

(2) Pyrimidinium compounds substituted in 2,5,6-position are obtained byreaction in 2-ethoxy ethanol under pressure in an autoclave at 100° C.with a reaction duration of 8 hours from the corresponding n-alkylbromides or iodides and the substituted pyrimidine compounds and theyields are between 30 and 40%. The recrystallizations are carried outfrom chloroform for all the substituted pyrimidinium compounds.

Preparative separation of unconverted products can be carried out asdescribed above by high pressure liquid chromatography.

(3) N₁ -n-alkyl compounds of pyrimidine can be obtained in good yieldsby reaction of n-alkyl-Mgx(x=Br, Cl) with pyrimidine or2,6,5,6-substituted pyrimidines in the presence of 1,2-dimethoxyethaneand/or n-heptane. No hetarine or addition-elimination orelimination-addition mechanism takes place.

0.01 mol (1.0 g) 5-fluoropyrimidine are dissolved in1,2-dimethoxymethane (100 ml) whilst stirring in a three neck flaskunder nitrogen. From a dropping funnel 0.08 mol (same order of magnitudeas above) n-decylmagnesium chloride (0.09 mo=29.6 g n-hexadecylmagnesiumbromide) dissolved in 20 ml heptane is added dropwise slowly at 20° C.

This solution is brought to 40° C, stirred for 12 hours and when thereaction is completed from a dropping funnel 20 ml 50% by weighthydrobromic acid is added dropwise at constant temperature. After 1 hourthe excess Grignard reagent is reacted. It is cooled to 0° C. and anyexcess of

Grignard reagent still present eliminated by adding methanol, thequaternary N₁ -pyrimidinium bases then being extracted by2-ethoxyethanol. The first recrystallization is carried out fromchloroform/methanol at 0° C. and the further recrystallizations at roomtemperature.

Melting point: 5-fluoro-N₁ -decylpyrimidinium bromide 199° C.decomposition)

Melting point: 5-fluoro-hexadecylpyrimidinium bromide 175° C.(decomposition)

(c) Preparation of 7 n-alkyl-imidazolium [4,5-d] pyrimidine derivatives(purine), e.g. 7-hexadecylimidazolium-2,6-dihydroxy[4,5-d] pyrimidinebromide 1.5 g 2,6-dihydroxy purine (0.01 mol are dissolved in 100 mlacetone in a four neck flask at 35° . From two dropping funnels whilststirring under nitrogen firstly triethyloxonium boron fluoride (Et₃O.sup.⊕ BF₄) in the excess (5.7 g=0.03 mol) with respect to n-hexadecylbromide (3.3 g, 0.01 mol) which is disposed in the second droppingfunnel is added dropwise simultaneously with n-hexadecyl Br. Thereaction is continued with constant stirring for 6 hours at 40° C. andsubsequently refluxing is carried out at 65° C. for 10 hours. Aftercompletion of the reaction 100 ml ethanol is added, the quaternaryammonium base formed filtered over a sintered-glass crucible (lG4) andrecrystallized from a mixture consisting of 2-ethoxyethanol/chloroform,1:1. Yield: 0.5 g, melting point: 122° C.

The compound is hygroscopic and forms a crystalline adduct with twoparts chloroform.

The UV spectra exhibit the typical absorption properties of the purinederivatives. This also applies to the ¹ H NMR spectra, measured in d₆-Me₂ SO₄.

(d) The corresponding benzothiazole and benzimidazole-n-alkyl compounds,particularly when they are halogenated in the 2-position, form with thisprocess with a yield of 50% and can be very easily recrystallized fromchloroform.

(e) The corresponding quaternary salts of the pyrazole may also beprepared by process (c). Process (b3) may also be employed withn-hexylmagnesium bromide or n-alkly-magnesium chloride because neitheran addition-elimination nor an elimination addition mechanism takesplace. The 4-H-pyrazolium salts with R=CH₃, OH, H form with a high yieldof 60%.

Since the n-alkyl radical can be localized both at the N₁ and at the N₂or both, the reaction product must be separated as described above byhigh-pressure liquid chromatograph in an RP-18 column in anacetone/acetonitrile elution mixture. This is also necessary when thecorresponding n-alkyl bromide is brought to react in a sealed tube orautoclave with a pyrazole derivative at 100° C. in the presence ofpiperidine. The ratio of di-N-substituted to mono-N₂ -substitutcdpyrazolium derivatives is 1.5:1.

(f) The imidazolium compounds, both the N₁ -substituted and the N₁, N₂-disubstituted, can be prepared like the corresponding pyridiniumcompounds.

To prepare the N₁ -substituted imidazolium compounds the proceduredescribed under (b3) is adopted. The yields are 30%. Acetone is asuitable reaction medium. (g) The quaternization of the pyrazine at theN₄ when substituted in the 2-position takes place with a 50% yield whenfor example a chlorine or a carboxamide (carbamoyl) group is located inthe 2-position. If the method under (b1) is adopted yields of 20-30% areobtained, depending on the size of the alkyl radical. If the knownprocedure for preparing pyridinium compounds (as adopted the yields areincreased to 50%.

As usual and as explained above the ((CH₂)_(x) chain with x=10-20governs the size and the cmc in aqueous solutions. The resulting size,form and molecular weight distribution of the micelle in aqueoussolution at pH 7.0 depend on the nature of the counter ion Y.sup.⊖.

The covalently bound pharmaceutical active substances may for example beextended to 9-β-arabino-1,4-adenine, 5-fluorocytosine, aza-uridine,6-mercaptopurine or thioguanine. These also include the nucleosides ornucleotides of the thymidine series which inhibit the growth ofneoplastic tumors inter alia by inhibiting the DNA synthasis. Also to bementioned here are the antiviral substances of the 1,3,5-triazines, e.g.the 2-acetamido-4-morphino-1,3,5-triazine, which has virustaticproperties against Herpes zoster.

                                      TABLE 2                                     __________________________________________________________________________    Characteristic properties of the N.sup.⊕ tensides of the general          formula HET N.sup.⊕ ═(CH.sub.2).sub.x --CH.sub.3 Y.sup.⊖                                   Analysis (%) found                               Nr.                                                                              HET N.sup.⊕ .tbd.(CH.sub.2).sub.x --CH.sub.3                                              Y.sup.⊖                                                                    Fp °C.                                                                      C  H  N  Y  cmc × 10.sup.-6 M              __________________________________________________________________________     1.                                                                              Hexadecyl-4-Hydroxy-                                                                          Br.1/2H.sub.2 O                                                                     85  59,86                                                                            10,94                                                                            4,37                                                                             24,83                                                                            0,95                                    pyridinium                                                                  2.                                                                              Dodecyl-pyridinium                                                                            Cl.H.sub.2 O                                                                        73  71,46                                                                            11,20                                                                            4,90  1,52                                  3.                                                                              2-Hydroxy,6-amino-hexadecyl-                                                                  Cl   155  61,45                                                                            18,69                                                                            10,76                                                                             9,10                                                                            2,00                                    pyrimidinium                                                                4.                                                                              Hexadecyl-pyrimidinium                                                                        Br   105  62,02                                                                            10,09                                                                            7,25  2,50                                  5.                                                                              2,6-Dihydroxy,5-Fluor,                                                                        Cl.2H.sub.2 O                                                                      172  61,13                                                                            22,68                                                                            7,14                                                                              9,05                                                                            0,85                                    hexadecyl-pyrimidinium                                                      6.                                                                              2-Hydroxy,5-methyl,6-amino-                                                                   Br   192  56,18                                                                            16,67                                                                            9,36  1,00                                    hexadecyl-pyrimidinium                                                      7.                                                                              Dodecyl-pyrimidinium                                                                          Cl    85  64,79                                                                            13,78                                                                            9,45  1,20                                  8.                                                                              2,6-Dihydroxy-dodecyl-                                                                        Br    70  53,07                                                                            17,12                                                                            7,75                                                                             22,06                                                                            1,90                                    pyrimidinium                                                                9.                                                                              2-Carboxamide-4-hexadecyl-                                                                    Cl   195(dec.)                                                                          71,30                                                                             6,77                                                                            11,89 0,30                                    1,4-diazinium                                                              10.                                                                              7-Hexadecylimidazolium-2,6-                                                                   Cl.1H.sub.2 O                                                                      112  60,80                                                                            17,13                                                                            13,51 0,50                                    dihydroxy[4,5,-d]pyrimidin                                                    7-Hexadecylimidazolium-2,6-                                                                   Br.1/2H.sub.2 O                                                                    170(dec.)                                                                          55,17                                                                             8,93                                                                            18,41                                                                            17,49                                                                            1,30                                    diamino-[4,5-d]pyrimidine                                                     3-Hexadecylbenzimidazolium                                                                    Cl.H.sub.2 O                                                                       100  72,53                                                                            10,80                                                                            7,35  6,70                                    4-Methyl- -2-hexadecyl-                                                                       Cl   172  69,67                                                                            11,89                                                                            8,14  0.70                                    pyrazolium                                                                    5-Methyl-1-hexadecylimidazolium                                                               Cl   142  69,69                                                                            11,89                                                                            8,12  3,90                                    3-Hexadecylthiazolium                                                                         Br.2H.sub.2 O                                                                      155  58,20                                                                            17,83                                                                            3,59  0,91                                    2,5-Dimethyl-3-hexadecyl-                                                                     Br.1H.sub.2 O                                                                      170(dec.)                                                                          57,15                                                                            20,50                                                                            3,34                                                                             19,01                                                                            15,00                                   thiazolium                                                                    3-Hexadecyl-6-methyl-benzimid-                                                                Cl.2H.sub.2 O                                                                      119(dec.)                                                                          69,81                                                                            14,13                                                                            7,09  17,00                                   azolium                                                                       3-Dodecyl-6-methyl-benzimid-                                                                  Br.1H.sub.2 O                                                                       98  59,40                                                                            12,52                                                                            7,29  7,30                                    azolium                                                                       3-Hexadecyl-5,6-dihydroxy-                                                                    Cl.2H.sub.2 O                                                                       70  60,60                                                                            28,54                                                                            3,07                                                                              7,79                                                                            7,90                                    benzthiazolium                                                             20.                                                                              3-Dodecyl-benzthiazolium                                                                      Br.1H.sub.2 O                                                                       90  70,20                                                                            14,57                                                                            4,31  10,90                                __________________________________________________________________________

                                      TABLE 3                                     __________________________________________________________________________    Yields and hydrodynamic radius of N-tensides of the formula                   HET═N--(CH.sub.2).sub.x --CH and benzethonium                             derivatives in dependence upon Y.sup.⊖                                                    Counter ion                                                                          <R.sub.H >                                                                        Yield                                          No.                                                                              Tenside          Y.sup.⊖                                                                      (Å)                                                                           (%)                                            __________________________________________________________________________    1  N-Cetyl-4-methyl-                                                                              Br.sup.⊖                                                                     140,0                                                 imidazolinium    Cl.sup.⊖                                                                      70,0                                                                             60                                                                 NO.sub.3 .sup.-                                                                       20,0                                                                             70                                             2  N-Hexadecyl-4-cetyl-                                                                           CL.sup.⊖                                                                     100 40                                                imidazolinium    HSO.sub.4 .sup.-                                                                     150 30                                             3  N-Hexadecyl-5-carboxamide                                                                      Br.sup.⊖                                                                     120,0                                                                             60                                                pyridinium       Cl.sup.⊖                                                                      55,0                                                                             60                                                                 Fumarate                                                                              70,0                                                                             70                                                                 Maleate                                                                              120,0                                                                             30                                             4  8-Ketohexadecylpyridinium                                                                      Cl.sup.⊖                                                                      50,5                                                                             00                                                                 Br.sup.⊖                                                                     140,0                                                                             80                                                                 NO.sub.3 .sup.-                                                                      170,0                                                                             100                                            5  Methyl-3-stearyloxy-propyl-                                                                    Cl.sup.⊖                                                                     140,0                                                                             60                                                pyridinium       Salicylate                                                                           1000,0                                                                            60-80 (20,25° C.)                       6  Cetyl-2,3-dihydroxy-propyl-                                                                    Cl.sup.⊖                                                                     150,0                                                                             20                                                hexadecyl-pyridinium                                                                           Br.sup.⊖                                                                     180,4                                                                             25                                                                 OH.sup.⊖                                                                     210,4                                                                             30                                                                 Maleate                                                                              120,0                                                                             41                                             7  3,5-bis[(n-hexadecyloxy-carbonyl]-                                                             Salicylate                                                                           1000                                                                              60                                                N-methyl-pyridinium                                                                            Fumarate                                                                             2500                                                                              70                                                                 Cl.sup.⊖                                                                     350 50                                             8  (a) 2,-4-Dihydroxy-5-methyl-                                                                   Cl.sup.⊖                                                                     1000                                                                              30                                                hexadecyl-pyridinium                                                                           Br.sup.⊖                                                                     1500                                                                              30                                                (b) 2,-4-Dihydroxy-5-Fluoro                                                                    Br.sup.⊖                                                                     210 30                                                hexadecyl-pyridinium                                                                           Cl.sup.⊖                                                                     150 30                                             9  (a) 2-Carboxamid-3-hexadecyl-                                                                  Cl.sup.⊖                                                                     220 30                                                1,4-pyridinium   NO.sub.3 .sup.-                                                                      440 30                                                (b) 2-carboxamid-3-dodecyl-                                                                    NO.sub.3 .sup.-                                                                      366 30                                                1,4-pyridinium   Fumarate                                                                             750 30                                             10 3-[[(Dimethylamino)-carboxyl]                                                                  Cl.sup.⊖                                                                     450 30                                                oxyl]-1-hexadecyl-pyridinium                                                                   Fumarate                                                                             700 60                                                                 Br.sup.⊖                                                                     1000                                                                              40                                             11 3-hexadecyl-benzimidazo-                                                                       Cl.sup.⊖                                                                     300 50                                                linium           Maleate                                                                              1500                                                                              40                                                                 Fumarate                                                                             250 30                                                                 NO.sub.3 .sup.-                                                                      500 70                                                                 SO.sub.4 .sup.2                                                                      350 70                                             12 Benzyldimethyl[2-[2-(p-1,1,3,3,                                                                Cl.sup.⊖                                                                     150 30                                                tetramethylbutyl-p,p' -dimethyl-                                                               Br.sup.⊖                                                                     3000                                                                              40                                                phenoxy)ethoxy]ethyl]ammonium                                                                  NO.sub.3 .sup.⊖                                                              150 10                                                                 Maleate                                                                              3000                                                                              20                                                                 Fumarate                                                                             2500                                                                              25                                                                 Salicylate                                                                           3000                                                                              20                                             __________________________________________________________________________

The following FIG. 4 shows the variance of the hydrodynamic radius ofbenzethonium chloride and N hexadecyl-4-cetyl imidazolium salicylate independence upon the hydrodynamic radius after various periods ofultrasonic treatment in minutes, measured by inelastic laser lightscattering.

Further preferred embodiments of the invention:

Whereas the overall range of the critical micellization concentration(cmc) is from 1.0 . 10⁻⁷ to 1.5 . 10⁻⁴ mol/liter, the cmc preferablylies in the range from 1.0 to 8.5 . 10⁻⁷ /liter.

Preferably, the cationic tenside with the monovalent anion is containedin ar amount of 0.05 to 0.1% by weight with respect to the totalpharmaceutical preparation. Particularly good results are achieved whenthe cationic tenside with the monovalent anion is contained in an amountof 0.08-0.1% by weight with respect to the total pharmaceuticalpreparation.

Preferably, the hydrophobic pharmaceutical active substance is containedin an amount of 0.06-0.5% by weight with respect to the totalpharmaceutical preparation.

Particularly good results are achieved when the hydrophobicpharmaceutical active substance is contained in an amount of0.001-0.005% by weight with respect to the total pharmaceuticalpreparation.

Preferably, the solvents are water or water+glycerol orwater+glycerol+ethanol. Preferably, the monovalent anion is a monobasicor dibasic fatty acid radical. Preferably, the monovalent anion isacetate, propionate, fumarate, maleate, succinate, aspartate orglutamate. Preferably the monovalent anion is a sugar radical.Preferably, the monovalent anion is gluconate, galacturonate oralginate. Preferably, the monovalent anion is chloride, bromide, odideor hydrogen sulfate. Preferably, the hydrophobic pharmaceutical activesubstance is an antimicrobial active substance or an antifungal activesubstance or an antiproliferative active substance or an antiviralactive substance.

Preferably, the hydrophobic pharmaceutical active substance is aninorganic compound of the elements zinc or mercury or tungsten and/orantimony. Preferably, the inorganic compound is Z_(n) SO₄ or Z_(n) O orHg(CN)₂ or (NH₄)₁₈ (NaW₂₁ Sb₉ O₈₆)₁₇ or an alkali or alkaline earth saltof phosphonic acid ROP(O)Me₂ or an N phosphonoacetyl-1-aspartate.

Preferably, the hydrophobic pharmaceutical active substance is anantibiotical and antiviral active substance or an antifungal activesubstance or an antineoplastic active substance.

Preferably, the solvent is water and/or ethanol and/or glycerol.Preferably, the solvent is water and/or ethanol and/ordimethylsulfoxide.

Whereas the pH value of the solvent must be ≦7, the preferable pH valueof the solvent=5 or is in the vicinity of 5.

The pharmaceutical preparation may be made according to the inventionsubstantially in that firstly the solvent is placed into a reactionvessel, then the cationic tenside is added whilst stirring at roomtemperature, then the hydrophobic pharmaceutical active substance isadded to the resulting isotropic micellar solution at room temperatureand stirring continued until complete dissolving thereof.

Particularly favourable results are achieved with the cationic tensidesof the general formula II when x=14, i.e. the alkyl chain has 15 Catoms.

These straight-chain C₁₅ derivatives of the N-tensides are distinguishedin particular by their simple chemical preparation. In addition, theysurprisingly have the lowest cmc (it is about 2.5.10⁻⁷ mol/liter). Theyare furthermore very easy to control by Y⁻ (form, molecular weightdistribution, polydispersity). Also, they are variable due to the sizeof the alkyl chain and thus as regards absorption of the pharmaceuticalactive substances. Finally, they can be easily crystallized.

As already mentioned the radical hexadecylpyridinium is known per se (aspure chemical compound) Not known is the influence according to theinvention of the associated anion (Y⁻) on the micelle size and the formof the micelle. With regard to the independent substance protectionclaimed according to the application for all the novel compoundsdisclosed a generic designation is given below which covers preferablythe novel compounds according to the invention. This term reads"isoelectronic heterocyclic nitrogen bases with 5 or 6 rings containingeither 2 N-atoms in the 1,2-position or 1,3-position or 1,4-position oran S-atom in 1-position with an N-atom in 3-position".

Production process for the pharmaceutical preparation General remarks onthe preparation of the aqueous phase

To obtain preferably a monodisperse homogeneous and isotropic aqueoussolution of the N⁺ -tensides both as regards form (spherical, oval,elongated) and size and as regards molecular weight distribution, thesolutions indicated, together with their included hydrophobicpharmaceutical active substances, must be

a. ultrasonically treated for example at 100 watt for one minute,possibly thereafter then by b,

b. subsequently purified by column chromatography, e.g. on an Agarose A0.5 m, Sepharose 2 B, Sephadex G 200, DEAE-Sepharose Cl-6B at pH 6.0 oran Ultragel AcA44 (pH 6.0-6.5) or BiO-Gel 1.5 m at pH ≦7.0; or

c. centrifuge on a linear density gradient, e.g. of 1-30% by weightsucrose, in a preparative ultracentrifuge in an SW-27 rotor at 25000 rpmfor 12 hours. When using a zonal centrifugation with the same gradient(20° C.) at 10000 rpm large amounts of homogeneous populations ofmicelles and vesicles can be centrifuged.

d. Purified by DEAE-Cellulose column chromatography at pH 5.0-6.5 (pH≦7), e.g. by phosphate gradient (linear from 0.01M KH₂ PO₄ /0.01M K₂HPO₄, pH 6.5 up to 0.05M KH₂ PO₄ /0.05M K₂ HPO₄ in the total elutionvolume of 1000 ml) until the desired population of micelles or vesicleshas been obtained.

It is thus possible to obtain the desired homogeneous populations ofmicelles or vesicles along with their included pharmaceutical activesubstances in the form of reproducible constant molecular weights andgeometrical configurations. This makes it possible to separatequantitatively monomers of the tensides from the micelles and fromunincluded pharmaceutical active substances.

Preparation of the homogeneous micellar solution in aqueous phase

The aqueous phase may be pure water. As a rule, however, an aqueoussolution cf an electrolyte is used. For example, an aqueous solution ofNaCl or CaCl₂ (MgCl₂) may be used. In addition, active pharmaceuticalagents of the aforementioned type may be introduced and are thendissolved in micellar manner, possibly subjecting the to sonicradiation.

Most processes are restricted to an encapsulation of hydrophilicsubstances. It is possible with the present invention to include inmicelles hydrophobic, for example lipophilic, inorganic (((Hg)CN)₂) andorganic active substances (amphotericin B). Also, hydrophilic anions ofpharmaceutical significance, for example salicylate, can be included atthe external surface of the micelle depending upon the nature of theN-tenside (in particular of formula II).

The invention can be employed to include either hydrophilic orlipophilic substances or both substances. In the case of hydrophobicactive substances the latter are then dissolved with the N-tenside ofthe formula I and II in a glycerol/ethanol mixture consisting of 15% byweight glycerol, 15% by weight ethanol and 70% by weight water or 50% byweight ethanol and 50% by weight water, possibly shaken orultrasonically treated and thereafter diluted to the aqueous phase witha content of glycerol/ethanol of at the most 15 g glycerol, 5 g ethanolin 100 g water. Subsequent gel permeation chromatography or preparativeHPLC can remove undesirable material and provide a homogeneous isotropicsolution. Whereas hydrophobic substances are made mainly via an organicphase (50%) and subsequent dilution (water), hydrophilic pharmaceuticalactive substances are preferably used in the aqueous liquid employed fordispersing the micellar solution. If necessary any unaccepted activesubstances can be removed from the dispersion using known techniques,e.g. dialysis, centrifuging, gel permeation chromatography.

The form and size and the degree of hydration of the micellar solutionsof the N-tensides depends inter alia on y⁻ and to a lesser extent on thestructure of the heterocycle although no doubt also on the hydrophobicchain length (CH₂)_(x). Thus, for example, in the presence of Br⁻ orsalicylate⁻ large rod-shaped micelles of hexadecylpyridinium can beobtained of an order of magnitude of L=10000 Å and a diameter of 100-500Å whereas in the presence of chloride micelles of the order of magnitudeof 50-100 Å are obtained in aqueous solution. In this case the shape andsize of the micelle defines the concentration of the (micellar) activesubstance to be encapsulated and thus behaves in a manner opposite toliposomes.

The advantage of the invention compared with the encapsulation withliposomes resides in

1. the density of these N-tensides which due to the previouslyaforementioned forces cannot liberate the micellarly boundpharmaceutical active substance and

2. the control of the form and size of the micelles by y⁻ and thus thecontrol of the absorptive capacity for hydrophobic and hydrophilicactive substances without major incisive influence of the heterocyclicon the cmc.

The resulting formation of the small and large micelles of theN-tensides in aqueous phase can be proved by physical measuring methods,e.g. with freeze-dried samples ("freeze fracture") under anelectronmicroscope or by X-ray small angle scattering, dynamic lightscattering, nuclear resonance spectroscopy (¹ H, ¹³ C and ³¹ P) and bytransmission electronmicroscopy.

In the nuclear resonance spectrum sharp signals with weak line width areobtained providing an indication of the formation of micelles with adiameter less than 600 Å. Sharp signals at δ about 0.89 ppm (--CH₃), δabout 1.28 ppm (--CH₂ --) and δ about 3.23 ppm (--N--(CH₃))₂ are forexample characteristic of the micelles of the N-tensides of the generalformula I and II. For included active materials in these micelles of theN-tensides a methyl signal at δ about 0.87 to 0.89 ppm is characteristicout is split into a triplet and has a substantially smaller line widththan the methyl signal which occurs as a singlet at δ=0.89 ppm but whichoriginates however only from the micelle.

These aqueous phases containing the micelles according to the inventionwith included active substances are administration systems whichpossibly after concentration, e.g. by ultrafiltration,ultracentrifugation or lyophilization with subsequent dissolving in anaqueous phase, are suitable for oral (p.o.) or local administration.

In the case of oral administration the micellarly bound pharmaceuticalactive substances of the N-tensides of the aqueous phase are mixed withpharmaceutically neutral diluents or carriers or with usual additives,for example coloring agents or flavouring agents, and administered assyrup or in the form of capsules.

Thus, a homogeneous isotropic micellar aqueous solution consistspreferably of an N-tenside of the formula II and I with an antiviralactive substance, in particular Hg(CN)₂, or ZnSO₄, ZnEDTA, idoxuridine,5-ethyl-2'-deoxyuridine or trifluorothymidine, amantadine, rimantadine(α-methyladamantane) and viderabine (9-β-arabino <1,4>-adenine) andribavirin (1-β-D-ribofuranosyl-1,2,4-triazole-3-carboxamide) and with2,6-di-amini-cubane 1,1':3,3'-bis-cyclobutane or singly substituted2,6-di-amino compounds (CF₃, Cl, OCH₃) dispersed in the presence orabsence of glycerol/ethanol (20° C.; ionic strength <0.2M).

A homogeneous isotropic micellar aqueous solution exists of an N-tensideof the formula II and/or formula I preferably with an antifungal activeagent, preferably with 5-fluorocytosine, clotrimazole, econazole,miconazole or oxyconazole (Z form) and with amphotericin B, nystatin andZnO.EDTA as inorganic antifungal active substance, and Hg₂ (CH)₄ Hg(CN)₂is present here as polymer, the dimer being the basic structural unit(dispersed in aqueous solution).

A homogeneous isotropic aqueous solution consists of an N tenside of theformula I and/or of the formula II preferably with an antineoplasticactive agent, in particular 5-fluorocyanide, Hg(CN)₂.4 (ascorbate oracetylacetonate), azauridine, cytarabine, azaribine, 6-mercaptopurine,deoxycoformycine, azathioprine, thioguanine, vinblastine, vincristine,daunoruticine, doxorubicine dispersed in the presence or absence ofglycerol/ethanol.

A homogeneous isotropic aqueous solution consists of an N-tenside mainlyof the formula II or the formula I preferably with amino glycosides suchas canamycin, gentamycin, neomycin etc. or tetracyclines,chloramphenicol or erythromycin as bacteriostatic (grampositive) orclindamycin (against nonsporiferous anaerobic bacteria) or rifampicin asbactericidal substance, and bacitracin, tyrotricin and polymycins,dispersed in the presence or absence of glycerol/ethanol.

The homogeneous mixture can also be subsequently dispersed in gels onthe basis of alginate, hydrogel structures such as Sephadex agarose,propyl cellulose, propylhydroxy cellulose, in the presence of DMSO,glycerol, the pharmaceutical active agents being contained micellarly inthe desired concentrations.

Dispersing is effected for example by vibration, stirring or ultrasonictreatment of the aqueous phase containing the previously madehomogeneous isotropic mixture. The formation of the micellar structureswith the included active substances, pH ≦7.0, 20° C., takes placespontaneously, i.e. without appreciable additional energy supply fromoutside, and at a high rate. The concentration of N-tenside of theformula I and II and the included compound can be increased if the cmcis exceeded by at least tenfold in the aqueous phase at constantchemical potential and temperature.

The cmc is a variable quantity for the amount of the monomers of theN-tensides which can be dissolved in a specific volume of wateremploying pH fluctuations ≦7.0. The cmc, which according to theinvention does not depend very much on the nature of the counter ion,which only governs the form, since the operation is carried out farabove the cmc, can be determined by electrochemical methods(conductivity, potentiometry) by measuring the transfer cells inconjunction with the counter ions, the surface tension, vapor pressurereduction, freezing point reduction and osmotic pressure, measuring thedensity, refractive index, the elastic and inelastic light scattering(diffusion coefficients, Stokes radius) and the viscosity, and bygelfiltration and X-ray small angle scattering measurements. Nanosecondsfluorescence and the measurement of the fluorescence polarization permitadditionally determinations of the pharmaceutical active substancesincluded by the N-tensides of the formula I and II, for example byZnEDTA or Hg(CN)₂ as quenchers and amphotericin B as intensifier.Positronium elimination measurements on these micellar solutionsdescribed with the included active substances also allow information tobe gained on the amount (concentration) of the included pharmaceuticalactive substance in dependence upon the nature and concentration of y⁻.

Aqueous phases having a pH value >7.0 are centrifuged after thedispersion. The neutralization to pH ≦7.0 is necessary to prevent adestruction of the heterocycle in formula I and of the active substanceand/or the micelles under basic conditions. Physiologically common andcompatible acids are for example diluted aqueous mineral acids andhydrochloric acid, sulfuric acid or phosphoric acid or organic acid, forexample low alkane acids such as acetic acid or propionic acid. Usuallythe aqueous phases of the cationic N-tensides of the formula I and IIreact acid to neutral but they can be exactly set to pH values between 3and 7.0 by Soerensen buffers or organic inert buffers such as HEPES,MOPS or MES.

Preparation of the homogeneous micellar solution in nonaqueous phases

The choice of the respective solvents depends on the solubility of theparticular pharmaceutical active substance. Suitable solvents are forexample methylene chloride, chloroform, alcohols, e.g. methanol, ethanoland propanol, low alkane carboxylic acid esters (acetic ethyl ester),ether or mixtures of these solvents. After preparation of the micellarsolution and adding the pharmaceutical active substance, dissolved inthe organic solvent, said organic solvent is removed either by themethods (a)-(b) mentioned above or by blowing off with inert gas, e.g.helium or nitrogen.

EXAMPLE 1

10 mg hexadecylpyridinium chloride is dissolved in 100 ml of awater/ethanol mixture (85:15; ^(w) /w) at 25° C. whilst stirring and 10ml of glycerol added. The pH value should be 6.5 but can be set with HClto said value or to another pH value (=7.0). This solution is thencooled to 20°±0.01° C. and then subjected to an ultrasonic treatment(Bronson Sonifier, Mass., U.S.A.) for two minutes at 10 watt. Theformation of the micelles is determined by measuring the diffusioncoefficient by means of inelastic light scattering and the Stokes radius(R_(H)) then calculated by the equation ##EQU1## In the presence ofCl.sup.⊖ as Y.sup.⊖ it should not be greater than 50 Å and in thepresence of Br.sup.⊖ it should not be greater than 1000 Å. To formmicroemulsions of micelles of specific size a film-like residue obtainedby evaporating the aforementioned solution in a rotary evaporator isdispersed at room temperature (20° C.) in 1/10 of the original volume by10-minute vibrating. A slightly opalescent aqueous solution is obtained.For inclusion of a pharmaceutical active substance, e.g. 5-fluorouracil,cytarabine or idoxuridine; these substances, which are sparingly solublein water, can be introduced directly, i.e. in solid form or as aqueoussuspension. Thus, for example, trifluorurdine, 1,0-3,0 mg, is added at20° C. whilst stirring either as microemulsion (suspension) or directlyto the aqueous micellar solution of the quaternary ammonium base. Aquantitative dosing of the aforementioned nucleoside and adeninenucleoside compounds can be achieved also by dialysis:

The micellar solution of the aforementioned concentration (buffered,unbuffered, pH ≅6.0, ionic strength variable, T=293° K.) is introducedinto a dialysis hose (the company Servant or Pharmacia), sealed andunder constant stirring at room temperature dialyzed for 2 hours againsta set solution of pH ≦7.0 which contains the aforementioned pyridineor/and adenine nucleoside of specific concentration. The decrease in theextinction at 260 nm with the time of the dialysis permits a check ofthe micellar incorporation of the aforementioned active substances intothe hydrophobic core of the hexadecylpyridinium chloride (Tab. 1).

                  TABLE 1                                                         ______________________________________                                        (20° C., pH 5.5)                                                                Concentration                                                                       Trifluoro-                                                     Experi-                                                                             R.sub.H (Å)                                                                        uridine    Idoxuridine  Yield                                  ment  (±5.0 Å)                                                                        mg/100 ml  mg/100 ml    (%)                                    ______________________________________                                        1     45,0      5          7,5    95   95                                     2     45,0      7,5       10,5    95   98                                     3     50,5     10,0       12,5    94   98                                     4     60,0     12,0       15,0    96   98                                     5     60,0     15,0       17,0    96   97                                     6     65,0     17,0       20,0    96   96                                     7     71,5     20,0       21,5    100  98                                     8     75,0     25,0       23,0    100  100                                    9     75,0     30,0       24,0    100  100                                    10    78,0     50,0       30,0    100  100                                    ______________________________________                                    

The resulting formation of small micellar structures in theaforementioned solution can be detected in the NMR spectrum by thesignals δ=1.25 (methylene), δ=0.86 (methyl). By incorporation of theaforementioned pharmaceutical active substances, depending on thesaturation in the hydrophobic core, a displacement of δ=1.25 (methylene)rakes place but not δ=0.86 (methyl).

The size of the micelles can be determined easily by inelastic lightscattering according to formula (1) (Table 1). The size and the shapefor obtaining a homogeneous and monodisperse solution can also beachieved by HPLC chromatography, gelpermeation and agarosechromatography. FIG. 5 shows, by curve no. 1, the temperature dependenceof Stokes' radius of 8-ketohexadecylpyridinium chloride with micellarlyincluded Z-miconazole, and by curve no. 2, the temperature dependence ofStokes' radius of 8-ketohexadecylpyridinium chloride with micellarlyincluded Z-miconazole+Hg(CN)₂.

A concentration of the micelles thus made can be achieved by pressuredialysis by means of fiberglass cartridges of defined pore size. It isalso possible to achieve not only a defined concentration ofpharmaceutical active substance but also to keep constant the micellesize, aggregation rate, hydration (solvation) because no fusion of themicelles "intermicellar growth") occurs. This means that the number ofmicelles pro volume unit increases with their included pharmaceuticalactive substance (concentration of hydrodynamic particles with the samemolecular weight) but not the aggregation rate or the number of anymonomers present which are separated by ultrafiltration.

EXAMPLE 2

Analogously to example 1 per test 15 mg benzethonium chloride isdissolved in 153 g water/ethanol (85/15; ^(w) /w) at 25° C. whilststirring and 0.5 ml glycerol added. The pH value is normally between 4.8and 5.5. To obtain a clear non-opalescent solution the latter issubjected to an ultrasonic treatment at 25° C. for two minutes at 20watt. The formation of the micelles of defined size is completed aftercooling to 20° C. after five minutes. For incorporation of theaforementioned antiviral active substances, e.g. trifluorouridine,idoxuridine, the procedure given under example 1 can be adopted.

For inclusion of miconazole (Z form) the micellar solution thus made isdispersed in the presence of miconazole of specific concentration,subjected to ultrasonic treatment (2 minutes), then chromatographed overagarose, and the micelles can be eluted with the hydrophobicallyincluded Z-miconazole as uniform monodisperse peak. The size andconcentration of active substance can be determined by inelastic lightscattering and UV spectroscopy. FIG. 6 shows, by curve no. 1, thetemperature dependence of Stockes' radius of benzyldimethyl{2-[2-(p-1,1,3,3-tetramethylbutryl-p,p'-dimethyl-phenoxy)ethoxy]ethyl}-ammoniumchloride with micellarly included viderabine; and by curve no. 2, thetemperature dependence of Stokes' radius of benzyldimethyl{2-[2-(p-1,1,3,3-tetramethylbutyl-p,p'-dimethyl-phenoxy)ethoxy]ethyl}-ammoniumchloride with 5-trifluoro-thymedine.

Analogously to example 1 10 mg benzethonium chloride and a desiredconcentration of Z miconazole can be dissolved each in 5 ml of achloroform methanol (3:1 mixture, then concentrated by hollow fiberpressure dialysis and thereafter dispersed in water or a desired buffer.A clear aqueous solution is obtained which comprises micelles of theorder of magnitude of R_(H) =60-80 Å in the presence of Cl.sup.⊖ orR_(H) =100-1000 Å in the presence of salicylate with included activesubstance.

By addition of 1% (^(g) /g) alginate and/or 5% (^(g) /g)dimethylsulfoxide thixotropic gels can also be made with theaforementioned included active substances. By increasing thebenzethonium chloride concentration, along with the included activesubstances, up to 2% (^(g) /g) effective oils can also be prepared.

EXAMPLE 3

Analogously to examples 1 and 2 the counter ions Y.sup.⊖ =Cl.sup.⊖,Br.sup.⊖ etc. can be exchanged after preparation according to theprocess by ion exchange chromatography o DEAE Sephadex A 50 or DEAESepharose or by dialysis exchange for the respective or desired counterion Y.sup.⊖.

(a) An aqueous micellar solution made by example 1 and 2 is brought topH=7.0 with 0.01N NaOH (20° C.). This can be done either by titration ordialysis against 0.01N NaOH for 10 hours. Subsequently, dialysis iscarried out against 1N fumarate or maleate solution, for which the Nasalts of fumaric or maleic acid can be used. The dialysis is completedafter 12 hours. A loss of antiviral active substances mentioned abovedoes not occur.

(b) An aqueous micellar solution, pH 6.0, made by example 1 and 2 iseluted on a DEAE Sephadex A 50 (1.0×100 cm) column previously chargedwith a buffered (0.01M K₂ HPO₄ buffer) 0.1N salicylate solution with aflow rate of 10 ml/30 min (20° C.). The excess salicylate is removed bydialysis against a large excess water/ethanol/glycerol (90/5/5; ^(g) /g)from the column eluate. The DEAE Sephadex A 50 chromatography can alsobe carried out under pressure by the countercurrent method with the samesolvent system. With exchange chromatography (DEAE Sephadex A 50, DEAESepharose 2B, 5B, DEAE-Cellulose, spherical) a homogeneous peak isobtained which can be analyzed by the criteria shown in examples 1 and2. DEAE Sephadex and DEAE Sepharose have the advantage that considerablequantities of micellar quaternary ammonium bases can both be purifiedand examined for monodispersity.

EXAMPLE 4

Analogously to example 1 a micellar solution of hexadecylpyridiniumchloride is prepared with the following pharmaceutical activesubstances:

    ______________________________________                                        100 g solution contain:                                                       ______________________________________                                        hexadecylpyridinium chloride                                                                          0.10   g                                              atropine hydrochloride (±)                                                                         0.002  g                                              zinc II chloride        0.004  g                                              glycerol                10.0   g                                              ethanol                 4.894  g                                              water                   85.0   g                                              pH                      6.2                                                   ______________________________________                                    

This preparation has a hydrodynamic radius of 35.0±5.0 Å and anaggregation rate of N=35 for a molecular weight of the monomer ofhexadecylpyridinium chloride of 393.0. Each micelle of this diametercontains on an average 100 μg zinc and/or 50 μg atropine (-).

FIG. 7 shows the variance in the hydrodynamic radius R_(H) of thispreparation. In FIG. 7, the heavy solid line representsN-dodecyl-5-carboxamidepyridinium fumarate at pH 5.8, the light solidline represents N-dodecyl-5-carboxamidepridinium fumarate at pH 5.8 plusatropine-HCl, the light dashed line represents cetylpyridinium chlorideat pH 5.5 plus Hg(CN)₂ atropine-HCl, and the heavy dashed linerepresents cetylpyridinium chloride at pH 5.5. It also shows theseparation according to the invention of the racemate atropine into theoptical antipodes, e.g. hyocyamine (-). The micellar size distributionis not changed by ZnII chloride.

FIG. 8 shows the variance in the hydrodynamic radius R_(H) of theN-hexadecyl-4-methylpyridinium chloride (dashed line) andN-hexadecyl-4-methylpyridinium chloride+atropine HCL (solid line).

EXAMPLE 5

5 mg 4-(17-tritriacontyl)-N-methylpyridinium chloride and 1-2.0 mgamphotericin B is dissolved in 10 ml of a chloroform/methanol mixture(2:1) under nitrogen at 25° C. and this solution is evaporated in arotary evaporator. The film-like residue is shaken in 5 ml distilledwater for five to 10 minutes. This solution is thereafter subjected toultrasonic treatment for three minutes until it is no longer opalescent.Depending on the requirements, this solution can subsequently be broughtto the pH value of 5.5-6.5 by adding 0.5 ml of a five-times concentrateof phosphate-buffered isotonic saline solution.

The solution made in this manner is introduced into a stirredultrafiltration cell (e.g. Amicon®) which is provided in place of theultrafilter with a straight-pore filter having a pore diameter of 0.05μm, filtered in the absence of Me²⁺ ions (Me²⁺ =Ca²⁺,Mg²⁺) so that thevolume in the cell does not drop below 30 ml. This results in vesiclesof a uniform size of <50000 Å.

The shape, size and molecular weight distribution can be determined asin examples 1 and 2. The pyridinium amphiphile is prepared from thecorresponding iodides with silver chloride in 10% (^(v) /v)ethanol/water. The colorless crystals have an Fp=64° C. (recrystallizedfrom acetone) and crystallize with one molecule of water.

1 H-NMR (CDCl₃ /Me₄ Si): δ 0.93 (6H,t,J˜4 Hz), 1.28 (60 H,m), 2.8(1H,q,J<2 Hz, not resolved), 4.75 (3H,s), 7.7-9.5 (4H,m). An H₂O-dependent signal at δ4.4 is characteristic.

Anal. calc. for C₃₉ H₇₄ NCl.H₂ O (MW 610.50) C 76.72; H 2-55; Cl 5.81;found: C 76.53, H 12.43; Cl 5.78.

EXAMPLE 6

Analogously to example 5 10 mg of 3.5-bis[(n-hexadecylonxy)carbonyl]-N-methyl-pyridinium chloride (Fp=102.5°)with 2.0 mg amantidine or rimantidine is dissolved in 10 ml of anethanol/water mixture (1:2) under nitrogen at 20° C. After ultrasonictreatment (5 min., 20° C., 10 watt) the vesicles formed with theirincluded active substances amantidine or rimantadine can be separated ina Sepharose 2B by size and molecular weight to obtain a homodispersesolution of vesicles with small molecular polydispersity. In the ¹ H-NMRspectrum the clear signals of methylene (δ=1.28) and methyl protons(δ=0.86) can be seen.

These unilamellar vesicles formed in examples 5 and 6 can be renderedvisible under an electron microscope. For this purpose the vesicledispersion is first subjected to the freeze-fracture method. This canalso be done by negative straining by means of the two drop method onFormvar or carbon grids. It is additionally possible by these twotechniques to render visible an populations of vesicles.

The method of inelastic light scattering used under examples 1 and 2makes it possible to determine the form and size of these vesicles andtheir included pharmaceutical active substances (FIG. 9). Curve 1 inFIG. 9 represents the temperature dependence of Stokes' radius of3,5-bis[n-hexadecyloxy)carbonyl]-N-methyl-pyridinium chloride withlamellarly included amantadine, not ultrasonically treated. Curve 2represents the temperature dependence of Stokes' radius of3,5-bis[(n-hexadecyloxy)carbonyl]-N-methyl-pyridinium chloride withlamellarly included amantadine, ultrasonically treated. Curve 3represents the temperature dependence of Stockes' radius of3,5-bis[(n-hexadecyloxy)carbonyl]-N-methyl-pyridinium chloride withlamellarly included rimantadine, ultrasonically treated.

3.5-bis [(n-hexadecyloxy)carbony]-N-methylpyridinium chloride,Fp=102.0-102.5° acetone). ¹ H-NMR (CDCl₃ /Me₄ Si): 0.85 (6H,t,J 5 Hz),1.31(56H,m), 4.40(4H,t,J<7 Hz), 5.03(3H,s) 9.20(1H,t,J<2 Hz),10.00(2H,d,J<2 Hz).

Analyt. calc.: C₄₀ H₇₂ NO₄ Cl(MW 666.47):C 72.10, H 10.88, C 15.32;found: C 71.44, H 10.84, Cl 5.23

EXAMPLE 7

3 ml gentamycin is dissolved analagously to examples 1 and 2 or in oneof the tensides named in Table 3 of the quaternary ammonium bases in 1ml of chloroform/methanol mixture (3:1) and this solution evaporateduntil a thin film is formed. This film is then dispersed in 10 ml water.Subsequently, the solution can be set to the desired pH >3<6.5 withbuffer. A clear solution is obtained.

This clear solution contains depending on the tenside used according toTable 3 a monodisperse distribution of micelles charged with gentamycinin the desired order of magnitude and yield (FIG. 10). Curve 1 in FIG.10 represents the temperature dependence of Stockes' radius(hydrodynamic radius) of N-cetyl-4-methyl-imidazolium chloride withmicellarly included rimantadine measured by inelastic laser lightscattering. Curve 2 represents the temperature dependence of Stockes'radius of N-hexadecyl-5-carboxamide-chloride with micellarly included5-fluorocytosine. Curve 3 represents the temperature dependence ofStokes' radius of 2,4-dihydroxy-5-methyl-hexadecylpyridinium chloridewith micellarly included gentamycin.

EXAMPLE 8

A micellar solution of hexadecylpyridinium chloride (cetylpyridinium) isprepared analogously to example 1 (20° C.) and contains the followingactive substances:

    ______________________________________                                        100 g solution contain:                                                       ______________________________________                                        cetylpyridinium chloride                                                                             0.10    g                                              atropine hydrochloride (±)                                                                        0.004   g                                              mercury II cyanide     0.004   g                                              glycerol               10.892  g                                              ethanol                5.0     g                                              water                  84.0    g                                              pH, T = 293° K. 5.7                                                    ______________________________________                                    

This preparation has according to the invention a hydrodynamic radius of35.0×10.0 Å and an aggregation number, n, of 35 with a molecular weightof the monomer of cetylpyridinium chloride of 393.0. Each micelle ofthis diameter contains on an average 5 μg Hg(CN)₂ and/or ˜5.0 μgatropine (-) (FIG. 12). In FIG. 12, the light line representshexadecyl-pyridinium-chloride, 0.2M NaCl, pH 5.8, and the heavy linerepresents hexadecylpyridinium-chloride, 0.2M NaCl plus Hg(CN)₂, pH 5.8.

This preparation is a homogeneous solution which contains micelles ofthe order of magnitude of 30-50 Å (R_(H)). It inhibits the growth ofinfluenza A virus as shown by the following Table 6 (FIG. 11). Curve 1in FIG. 11 represents the temperature dependence of Stokes' radius ofN-hexadecylpyridinium chloride and micellarly included Hg(CN)₂, notultrasonically treated. Curve 2 represents the temperature dependence ofStokes' radius of N-hexadecylpyridinium chloride and micellarly includedHg(CN)₂, ultrasonically treated.

                  TABLE 6                                                         ______________________________________                                                       Titration of infection.sup.b,                                  Inhibitor.sup.a                                                                              Plaque forming units                                                                         Inhibition.sup.c                                ______________________________________                                        1-adamantaneamine HCl                                                                        2 × 10.sup. 6                                                                          -1.11                                           Aqueous Hg(CN).sub.2 solution                                                                1 × 10.sup. 6                                                                          -1.30                                           Cetylpyridinium chloride                                                                     1.5 × 10.sup. 8                                                                        -0.11                                           Preparation according to                                                                     2 × 10.sup. 5                                                                          -1.45                                           example 8                                                                     Check          2 × 10.sup. 8                                                                          --                                              ______________________________________                                         .sup.a Inhibitor concentrations are added in the in vitro cell cultures o     100 μM.                                                                    .sup.b The plaque assay was carried out in accordance with K. Tobita, A.      Suginire, C. Enamote and M. Fusiyama, Med. Microbiol. Immunol., 162, 9        (1975) on renal epithelial cells (dog, MDCK) in the presence of trypsin.      .sup.c The inhibition is given as the negative decadic logarithm of the       quotient of the plaque forming units in the presence of the inhibitor to      that without inhibitor: .sup.10 log (pfu/ml of the inhibitor/(pfu/ml          check).                                                                  

FIG. 7 shows the variance in the hydrodynamic radius R_(H), of thispreparation. It also shows the separation described above according tothe invention of the atropine into its optimum antipodes in the presenceof Hg(CN)₂.

EXAMPLE 9

5 mg of an N-quaternary ammonium base given in Table 3 (usually No. 1,2or 4) and 2.0 mg 5-fluorouracil or 1.5 mg 5-fluorodeoxyuridine isdissolved in 10 ml of a chloroform/methanol/ether mixture (3/1/1) andthis microemulsion dispersed by vigorous shaking at 25° C. for twohours. There are two methods for the further processing:

(a) The suspension is evaporated to form a thin film (under N₂ and UVprotection). The film-like residue is then dispersed in water or buffer,for example 0.01 M to 0.001 M KH₂ PO₄, set to pH 4.5-6.5. Afterpreviously subjecting this partially opalescent solution to ultrasonictreatment (10 watt 2 min) to increase the yield, the clear micellarsolution is subsequently separated on a Bonder Pack I-250 or an RP 18column by high-pressure liquid chromatography (HPLC) from any monomerspresent and any unincluded pharmaceutical active substances. In astirred ultrafiltration cell (Amicon®) concentration is carried out witha filter of polycarbonate with a pore diameter of 0.015 μm.

(b) 10% (^(g) /g) dimethylsulfoxide (DMSO) and 2.5% (^(g) /O) alginateare stirred into this suspension at 25° C. The resulting gel formsspontaneously. In the X-ray small angle diagram a uniform spacing ofd=125 Å is found which is very different from alginate gels (d=25.45 Å).The gel has thixotropic properties and becomes liquid at 45° C.Reformation of the gel takes place at 42° C. and its constantrheological parameters are achieved after 2 hours at 20° C. and 37° C.respectively.

The final concentrations per 100 g pharmaceutical preparation are asfollows:

    ______________________________________                                        100 g solution contain:                                                       ______________________________________                                        (a)     N.sup.+ -tenside (Table 3, No. 4)                                                                0.01 g                                                     5-fluorodeoxyuridine                                                                             0.10 g                                                     glycerol          11.89 g                                                     water             88.00 g                                                     T = 293° K., pH = 5.5                                          (b)     N.sup.+ -tenside (Table 3, No. 2)                                                                0.05 g                                                     5-fluorodeoxyuridine                                                                             0.05 g                                                     dimethylsulfoxide 10.00 g                                                     alginate           2.50 g                                                     water             86.50 g                                                     T = 293° K., pH = 5.5                                          ______________________________________                                    

EXAMPLE 10

15 mg (0.02 mMol) benzethonium chloride and 2 mg2-acetamido-4-morpholino-1,3,5-triacine are dissolved in 30 ml of awater/ethanol mixture (80:20 or 90:10) at 20° C. under ultrasonictreatment in 0.01 M K₂ HPO₄, pH 6.5, under an N₂ stream. An opalescentaqueous phase is obtained. By separating the reversed micelles from themicelles in aqueous phase on a Sepharose 2B column (1.5×100 cm) auniform monodisperse micelle formation is obtained having an averagehydrodynamic radius of 50 Å. The chromatograph solution can beconcentrated as in example 9 by an ultrafiltration. The solution isstabilized by using 5% (^(w) /w) glycerol or adding 2% (^(w) /w)salicylate. The solutions thus made do not change their hydrodynamicradius, their partially specific volume or molecular weight distributionin the temperature range of 15-45° C.

    ______________________________________                                        100 g solution contain:                                                       ______________________________________                                        benzethonium chloride  0.15    g                                              2-acetamido-4-morpholino-                                                                            0.006   g                                              1,3,5-triazine                                                                salicylic acid         0.05    g                                              glycerol               5.00    g                                              water                  94.894  g                                              T = 239° K., pH = 5.5                                                  ______________________________________                                    

EXAMPLE 11

30 mg (0.020 mMol) 3.5-bis [(n-hexadecyloxy)carbonyl]-N-methylpyridiniumchloride and 1.0 mg (˜0.005 mMol) polyoxin A are dissolved in 10 ml 0.01M KH₂ PO₄, pH 6.5, at 20° C. containing 1 ml of a mixture of tert.butanol/methanol/ethanol (2:1:1). The solution is ultrasonically treated(20 watt, 5 min) in an ice bath at 0° C. and thereafter made up to 20 mlwith phosphate buffer, pH 7.0. The clear non-opalescent solution ischromatographed on a Sepharose 2B column at pH 7.0 in the presence ofphosphate at room temperature The vesicles doped with the pharmaceuticalactive substance are concentrated in an ultrafiltration cell (Amicon®)with a pore diameter of 0.05 μm under slight excess pressure. Afterpassage of 0.3-0.5 ml filtrate al the vesicles with a diameter of 350Åare separated and the supernatant dispersion can be introduced intoampoules and used for therapeutic tests. FIG. 13 shows the inhibition ofchitin synthetase in digitonin-treated cells (Saccamyces cerivisiae andCandida albicans) after addition of this preparation in dependence uponthe polyoxin A concentration in FIG. 13, the points represented bysquares indicate data taken with polyoxine A, the points represented bytriangles indicate data taken with polyoxine A plus Hg(CN)₂ (D4), andthe points represented by circles indicate data taken with polyoxine Aplus ZnCl₂ (gluconate) (D4).

EXAMPLE 12

Analogously to example 2 10 mg Hg(CN)₂ and 40 mg Na ascorbate at pH 7.0are dissolved in 10 ml phosphate buffer. The suspension is subjected toan ultrasonic treatment at 0° C. for 5 min, slowly heated to 20° C. andcentrifuged to a 10% (^(w) /w) linear glycerol gradient in a preparativeultracentrifuge at 1000 xg for 6 hours (20° C., Polyalomer tubes). Afterthe dripping out the Uv-active fractions are united and concentrated inan Amicon flow cell and subsequently analyzed for Hg and ascorbate(HPLC; mobile solvent CH₃ OH/H₂ O (50/50) Hg detection withNa-diethylthiocarbamate, hexadecylpyridinium Cl, e.g. by Uv detection at251 nm; Hg (CN)₂ ascorbate by UV detection at R=245 nm at pH 2.0 andR=265 nm at pH 7.0). These micellarly included Hg(CN)₂ ascorbatecomplexes (MW-1.500) have in accordance with Table 5 the followingrepresentative inhibitor concentration: with respect to B. subtilis DNApolymerase III.

                  TABLE 5                                                         ______________________________________                                                                      Hg(CN).sub.2.                                                                         Competi-                                                     Hg(CN)   Ascorbat                                                                              tive                                    Nr.  Tensid          K.sub.i, μM.sup.2                                                                   K.sub.1, μM                                                                        with                                    ______________________________________                                        1    Hexadecyl-pyridinium-                                                                         7,9      15,3    dGTP                                         Cl.sup.⊖                                                         2    Hexadecyl-pyridinium-            dGTP                                         Cl.sup.⊖                                                         3    Benzethonium-Cl 33,1     12,0    dATP                                    4    Benzethonium-Cl                  dATP                                    5    8-Keto-hexadecyl-                                                                             0,4       0,05   dGTP                                         pyridinium-Cl                                                            6    8-Keto-hexadecyl-                                                                             2,5      7,5     dGTP                                         pyridinium-Cl                                                            7    3,5-bis (n-hexadecyloxy-                                                                      2,0      9,2     dGTP                                         carbonyl -N-methyl-                                                           pyridinium-Cl                                                            8    4-(17-tritriacontyl)-N-                                                                       4        10,1    dGTP                                         methyl-pyridinium-Cl                                                     9    acc. to Table 3 Nr. 9                                                                         9        0,5     dGTP                                    10   acc. to Table 3 Nr. 10                                                                        0,1      7,9     dATP                                    ______________________________________                                    

The inhibitor concentrations are given in 50% of the completeinhibition. The assay which was used is that according to Clements, J;D'Ambrosio, J; Brown, N. C; J.Biol.Chem. (1975) 250, 522 and Wright, G.E.; Brown, N. C; Biochem. Biophys. Acta (1976) 432, 37.

PHARMACODYNAMIC TESTS

The significance of highly reactive oxygen molecules (superoxideradicals O₂, peroxides H₂ O₂, hydroxyl radicals . OH, sing let oxygen ¹O₂) in the inflammatory process is known (cf. e.g. McCord, J. M., K.Wong; Phagocytcsis-produced free radicals: roles in cytotoxicity andinflammation. In: Oxygen Free Radicals and Tissue Damage, ExcepterMedica, Amsterdam-Oxford-New York, 1979, 343-360; Allgemeine undspezielle Pharmakologie, Herg. W. Forth, D. Henschler, W. Rummel,Biowissenschaftlicher Verlag, 1983). They arise inter alia in thephagocytosis by activated leucocytes (monocytes, macrophages,polymerphonuclear, neutrophilic granulocytes) and can be used forkilling exogenous cells and bacteria, bacilli, etc., and for certainviruses when the immunological system and the receptors of thephagocytes specific to IgG or the complementary component C₃ arefunctioning normally. The phagocytizing cells themselves areintracellularly protected from damage by these particularly active formsof oxygen by a system consisting of several enzyme systems.

It has now been found that quaternary ammonium bases of the generalformulae I and II ##STR25## wherein Y.sup.⊖ may be a counter ion both ofan inorganic, e.g. Cl.sup.Δ, Br.sup.⊖, H₂ PO₄ ⁻ or organic nature, e.g.fumarate, malate, salicylate, acetate, propionate, gluconate andalginate and the heterocycle may be both a pyridine, pyrimidine,pyrazine, imidazole, thiazole or purine, but a π-excess or π-defectivearomatic system, which are all able at pH ≦7.0 to eliminate these oxygenradicals in accordance with the following reaction mechanism: ##STR26##All reactions which take place in the inflammatory range between pH 5.0and 6.0 require a pH range ≦7.0, which is ensured by the preparationsmade according to this invention. The resulting aggressive oxygenradicals are intercepted in accordance with the reaction 1-4 by theN-tenside, e.g. cetyl-pyridinium chloride, as are the resulting hydratedshort-life electrons which can originate from collisions of O₂ ⁻radicals with H₂ O. As a result the N-tensides in the pH range ≦7.0according to the invention have a membrane-protective effect so that theinflammation reactions according to a prostaglandin mechanism cannotoccur. The high capture rate of .O₂ ⁻ radicals in the N-tensides ofk=5×10¹² M⁻¹ and its dependence on the ionic strength, which however canbe held constant by adding ethanol/glycerol, is explained by theelectrostatic double-layer structure of the quaternary ammonium bases.

Thus, the invention prevents misdirected lytic reactions in whichaggressive oxygen radicals participate as pathogenic mechanisms of theinflammatory diseases due to microorganisms and viruses. Thus, interalia the cytotoxic effect of the resultant products of these aggressive.O₂ ⁻ radicals is prevented by the N-tensides according to the inventionas shown by the example of cetylpyridinium halide, and inter alia theinvention prevents depolymerization of hyaluronic acids, proteoglycanes,collagen fibriles, cytoskeletons, etc., this also applying to mucous andmembranous tissues (outer surfaces).

Furthermore, with the preparations made according to the processdescribed it has been found that compounds of the structure I and IIreduce the infection of human cells in vitro so that the micellarsolutions I and II made according to the invention represent aprotection for the cells and their external surface.

It has further been found that this protection is intensified byincorporation of Hg(CN)₂, ZnEDTA and/or antiviral, anti fungal andantibacterial active substances.

It was found that on incubation of monolayer cell cultures, infectedwith influenza virus, subgroup A₂, of Vero cells and also with Herpessimplex virus HSV I-III in vitro more than 60% of the cells areprotected from infection by the respective virus.

It has further been found that the effect of the protection by the N⁺-tensides according to the general formula I and II for monolayer cellfunctions in vitro is not intensified by the antiviral active substancesalthough the inhibition concentrations of the antiviral activesubstances are lowered by 30% by cytarabire, idoxuridine,trifluorothymidine, as well as monolayer cells infected with Herpessimplex virus type 1 or influenza virus type A₂, compared withapplications not containing any quaternary ammonium bases according toformula I and II. The combination of N⁺ -tenside according to thegeneral formula I and II thus proved to be the effective virostatic incombination with micellarly bound antiviral active substances (FIG. 2).

It was further found that the N⁺ tensides according to the generalformula I and II intensify the antifungal effect in combination withantifungal active substances such as econazole, clotrimazole andmiconazole (≈35%) since the N⁺ base with a suitable counter ion is ableto extract cholesterol from the external membrane of the fungus orhyphae to form mixed micelles and is then able to inject the antifungusactive substances, which are again bound, into the cell interior of thefungus.

It has further beer found that the antifugal effect is intensifiedtenfold by a mechanism hitherto unknown by amphotericin B and N-tensideof the formula II, preferably hexadecylpyridinium bromide, decyl andhexadecyl-1-pyridinium chloride or hexadecyl-4-hydroxypyridiniumchloride. This means that in accordance with the invention asubstantially smaller active substance concentration of thepharmaceutical agent suffices to achieve the same therapeutical effects.

It has been found inter alia that the fungistatic effect is intensifiedby micellar incorporation of ZnEDTA and ZnO, in particular also byHg(CN)₂, into the N-tensides of the formula I and II, in particular inthe case of hexadecyl pyridinium chloride andhexadecyl-4-hydroxy-pyridinium bromide, in particular at concentrationsof the inorganic active agents at which said agents themselves are notyet effective.

It has been found that according to the invention the micelles of theN-tensides -n the aqueous phase at pH≦7.0 can micellarly bindtherapeutic amounts of benzoylperoxide, which is sparingly soluble inwater and alcohol. Thus, for example, 1 g benzoylperoxide can bedissolved in 20 ml benzethonium chloride or in 25 ml hexadecylpyridiniumchloride, in particular however in 3-hexadecylbenzothiazonium bromide.On local administration the micellar solution causes at the skin similarpeeling effects a Tetrinoin. Due to its additional very bacteriostaticproperties, both of the benzoylperoxide and of the N-tenside, thiscombination according to the invention is particularly suitable in thecase of inflammatory forms of acne, e.g. acne comedonica, acnepapulo-pustulosa and acne conglobata.

It has been found that the micelles made according to the invention inaqueous phase of the N-tensides, in which Hg(CN)₂ or ZnO, ZnSO₄, ZnEDTAis micellarly included, in the cell culture irreversibly andvirus-specifically inhibit the reproduction of Herpes simplex virusesdue to inhibition of the viral DNA polymerase. The non-infected cellsremain largely uninfluenced so that the methods according to theinvention described for example for hexadecylpyridinium chloride,3-hexadecylbenzothiazolium bromide (sulfate), including theaforementioned inorganic active substances, lead to a therapeutic agentinvolving no risk. The astringent properties of Hg(CN)₂, ZnO, ZnSO₄,ZnEDTA play no part because in the hydrophobic core of the micellesthere are no free ions since a) for example Hg(CN)₂ (more correctly Hg₂(CN)₄)) is undissociated, (b) the inorganic active substances areincluded by their lipophils and (c) there is practically no water in thehydrophobic core.

The combined effect, the formation of mixed micelles of the N-tensidesaccording to the general formula I and II with the membrane affected bythe virus and and the phospholipid double membrane of the virus itselfand the subsequent antiviral effect on the virus DNS polymerase by theaforementioned inorganic and organic active substances such as5-ethyl-2'-deoxyuridine and viderabine, analogous to the nucleoside, wasdetected as illustrated in FIGS. 2a, b.

It was also possible to detect this mechanism in the case of rhino andinfluenza viruses. Other effects were found, with however smaller activesubstance concentrations, for 1,1': 3,3'- biscyclobutane and for1,1':3,3' amine-substituted cubanes.

It was found that the intensified antiviral effect for phospholipidviruses, adeno viruses and Herpes simplex I due to the N-tenside and themicellarly included active substances develop their effectsynergistically by the following biochemical mechanisms:

(a) Binding to DNA, RNA-forming enzyme systems, unfolding of thepolypeptide chain is intensified by the N-tenside (denaturing).

(b) Template binding, e.g. daunomycin, adriamycin

(c) Binding of nucleoside analogs, e.g. the aforementionedara-CTP-C5'-triphosphate of cytosine arabinoside, azathioprine

(d) Binding of inorganic active substances, e.g. Z_(n) SO₄, Z_(n) O,Hg(CN)₂, wolframic acid antimonates, e.g (NH₄)₁₈ (NaW₂₁ Sb₉ O₈₆)₁₇ andK₁₈ (KW₂₁ Sb₉ O₈₆)₁₇, as well as Hg-substituted cubanes of theaforementioned type. In combination with the antiviral effect of themicellarly included antiviral active substances employing the procedureaccording to the invention a reduction of the ED₅₀ by 20-25% in vitrocompared with the pure active substance is noted so that the samemolecular biological action can be achieved with an approx. 20% dose bythe micellar effect. This applies in particular to micellarly includedrubaricine in hexadecylpyridinium bromide, hexadecylbenzothiazoliumchloride and benzethonium chloride. DNA viruses and Herpes viruses aremost sensitive in these examples, in contrast to rimantadine+N-tensidesof the formula I and II, which are primarily effective in vitro againstRNA viruses.

It has further beer found that the antitumor activity ofadenosine-desaminase inhibitors dissolved micellarly according toformula I and II by the process of the invention, e.g.erythro-9-(2-hydroxy-3-nonyl)-adenine, deoxycoformycin, is intensifiedtenfold. The same was found for aspartatetranscarbamylase inhibitors;thus, the biosynthesis of pyrimidine was intensified 20-fold bymicellarly included N-(phosphono acetyl)-aspartate by blocking of thecarbamylation of aspartate.

It has also been found that both micellar)y included Hg(CN)₂, ZnSO₄, ZnOor ZnEDTA, as also 5-trifluoromethyl 2'-deoxyuridine, which is formed invitro from trifluoromethyl uracil, irreversibly inhibits the thymidinesynthetase, a key enzyme of the DNA synthesis.

The pyrimidine biosynthesis is irreversiby inhibited by 20% bypyrazofurine, a naturally occurring antibiotic which is micellarlyincluded, and at the same time the cell toxicity is reduced.

It has further been found that the diffusion barriers for antibiotics,e.g. tetracyclines, aminoglycocides, this applying to β-lactamantibiotics (penicillin), after a certain time in the case of E. colibacteria are reduced for the micellarly included effective substances.This diffusion barrier is concentration-dependent for the aforementionedactive substances but not for the N-tensides prepared according to theinvention. These are folding processes at the outer membrane, primarilya change in the structure of the porines within the outer membrane of E.coli, so that for example the inorganic active substances Hg(CN)₂,ZnSO₄, ZnEDTA can diffuse into the periplasma of the external cellmembranes of gramnegative bacteria.

The porines are membranous water-filled pores through which thehydrophilic pharmaceutical active substances can diffuse into theinterior of the cell. Hydrophobic pharmaceutical active substancescannot pass through these porines. The N⁺ -tensides, in particular ofthe general formula HET.tbd.N-(CH₂)_(x) --CH₃ y.sup.⊖ and alsobenzethonium derivatives, can pass through these water-filled pores.Thus, micellarly included pharmaceutical hydrophobic (lipophilic) activesubstances, in particular of an inorganic nature, due to the hydrophilicouter form of the N⁺ -tensides can reach the cell interior by passivediffusion. There they then react also with the cell-wall-synthesizingenzymes, in particular in the case of Hg(CN)₂ at concentrations of 10μg/ml and ZnEDTA at c=5 μg/ml. The rate of the diffusion of micellarlyincluded active substances increases with increasing hydrophobiccharacter; normally, this is exactly converse, e.g. the diffusion ratefor gramnegative bacteria decreases with increasing hydrophobiccharacter. Furthermore, a positive charge promotes the diffusion and theformation of mixed micelles of these N-tensides to be prepared accordingto the invention. The validity of these findings could be proved as afunction of the concentration by investigations of the diffusion anddissolving rates of various radioactively (C¹⁴) marked N-tensides at themembrane (periplasma).

It was also found in vitro that the thymidilate synthetase (TS)(EC2.1.1.45) is inhibited both by Hg(CN)₂ in aqueous solution and inmicellar solution of an N-tenside of the formula I and II, in which theHg(CN)₂ is hydrophobically dissolved. TS catalyzes the conversion ofdUMP and CH₂ --H₄ folate to dTMP and H₂ folate. Since this enzyme isessential to the synthesis of dTMP, i.e. to DNA synthesis itself, itthus represents a target for pharmaceutical active substances againstneoplastic cells. It has now been found that for example a solution madeaccording to the invention of hexadecylpyridinium chloride which keepsHg(CN)₂ hydrophobically bound, has the cytostatic activities listed inTable 1 against leukemia cells (L1210 cells). It was thus possible tofind inter alia that TS, dUMP and Hg(CN)₂ as inorganic pharmaceuticalactive substances form a ternary complex according to (A, B) ##STR27##which can be isolated by column chromatography on Sephadex G-25 andBiO-Gel P10. The formation of the complex according to equation A has aformation constant of k₁ =0.51 h⁻¹ in the case of hexadecylpyridiniumchloride and k₁ =0.70 h⁻¹ in the case of benzethorium chloride andmicellarly included Hg(CN)₂. The dissociation constants are k₋₁ =0.015h⁻¹ (CPCl) and k₋₁ =0.02 h⁻¹ i.e both are very slow, that is theformation and the dissociation of the complex. In contrast the formationof the dimer according to B is substantially faster: k₁ =0.02 h⁻¹ andk₋₁ =0.015 h⁻¹ for CPCl and k₁ =0.01 h⁻¹, 0.03 h⁻¹ for benzethoniumchloride. This means that micellar solutions of quaternary ammoniumbases according to formua I and II at pH≦7.0 which keep Hg(CN)₂hydrophobically bound are therefore therapeutical for slowly growingtumors where other inhibitors are ineffective as regards TS and theobserved cytotoxicity for the normal cells of other antimetabolites cantherefore be slowed down in the case of rapidly growing proliferatingcells.

Ribavirin, which is a synthetic 1,2,4-triazol^(e) nucleoside, has abroad antiviral spectrum for DNA and RNA viruses. Ribavirin micellarlyincluded by cationic tensides of the form (HET=N⁺ --(CH₂)_(x)--CH₃)Y.sup.⊖ passes very rapidly through the membrane barrier, morerapidly than the pharmaceutical active substance itself. The conversionof ribavirin to monophosphates, diphosphatas and triphosphates is alsoincreased by the specific cellular enzymes so that the inhibition of thevirus-induced enzymes necessary for the viral nucleic acid biosynthesisis accelerated. Whereas ribavirin on its own only has a moderate effecton the cellular DNA synthesis and is cytotoxic in the range of 200-1000μg/ml for normal cell lines, the cytotoxicity drops in the presence ofcationic micelles, when it is micellarly included, to 50 μg/ml (in vitrotests), measured with respect to cells infected with Herpes simples (DNAvirus).

Amantadine (1-adamantanamine-HCl) has a particular pharmadynamic actionagainst influenza viruses (class A). The replication of most influenza Astrains is inhibited in vitro between 0.2-0.6 μg/ml. Micellarly includedamantadine in cationic micelles, in particular of the form(Het.tbd.N-(CH₂)_(x) --CH₃)Y.sup.⊖, effect a reduction of theconcentration of pharmaceutical active substance to 0.05 μg/mlamantadine, measured by Hayden et al. (Plaque inhibition assay for drugsusceptibility resting of influenza viruses. Antimicrob. Ag. Chemother.1980, 17: 8657-70; Grunert et al.; Sensitivity of influenza A/New Jersey18/76/(Hswl-Nl) virus to amantadine HCl, J. Inf. Dis. 1977, 136,297-300). Whereas amantadine has practically no activity againstinfluenza virus type B inhibition is exhibited by micellarly includedamantadine in the cationic tensides of the formula ##STR28## With aconcentration of 0.01% by weight of amantadine for influenza virus typeB corresponding to a concentration of 0.5 μg/ml pharmaceutical activesubstance in vitro.

A surprising effect of micellarly included amantadine in the twocationic tensides of the above formula has been found: whereas theinfluenza virus type A is not resistant to amantadine in vitro it isresistant with amantadine on its own.

Rimantadine-HCl (α-methyl-1-adamantanemethylamine hydrochloride) has thesame pharmacodynamic actions in vitro as amantadine but has a greatereffect for the same dose. Here too it has surprisingly been found thatrimantadine micellarly included in a cationic tenside, in particular ofthe two above formulae, has the same in vitro effect as the purepharmaceutical active substance although with a substantially smallerdose of 0.05 μg/ml.

Of the inorganic pharmaceutical active substances, Hg₂ (CN)₄ andmicellarly bound Hg(CN)₂ in cationic micelles exhibit a surprisingantiviral interferon-induced spectrum. In cell cultures of lymphocytesand fibroblasts it was possible to detect an induction of interferon α₁and interferon β after incubation with micellarly included Hg(CN)₂ in acationic tenside of the formula ##STR29## at a concentration of Hg(CN)₂of 5 μg/ml to 15 μg/ml in a 0.1% (^(g) /g) cationic tenside. In the caseof interferon α₁ concentrations of 20-50 units/ml were found and in thecase of interferon β 10-20 units/ml. The micellar incorporation ofmercury cyanide increases the liberation of the interferon α₁ inparticular however of the interferon β in the case fibroblast culturesby 10 to 100-fold compared with simple aqueous buffered solutions ofmercury II cyanides.

Secondary effects

The observed secondary effects of interferon α₁, for example headache,lymphocytopenia, slight to medium sickness symptoms, are not present, orwere not observed, with the pharmaceutical preparations described here,in particular against influenza and rhino viruses. This is due primarilyto the fact that substantially less units/ml of interferon A, induced bythe inorganic pharmaceutic active substance, are used therapeuticallythan in a therapy with interferon α on its own. Thus, no toxic effectsare observed, for example gastrointestinal disturbances, loss of weight,alopecia, peripheral spasms, paraesthesia and tone mark depressions,although these are reversible.

The hematological toxicity, which in the case of interferon α₁ dependson the dose (threshold dose 3×10⁶ units/ml), does not manifest itselfwith these pharmaceutical preparations for mercury cyanide, rimantadine,amantadine and ribavirin.

(b) The pharmaceutical preparation consisting for example ofhexadecylpyridinium chloride or a pyrimidinium derivative and ahexadecyl radical in the presence of micellarly included mercury cyanideresults in interferon production in the cell culture. This is aninterferon induction by liberated mercury cyanide which occurs locallyin high concentrations and with a relatively high molecular weight of4500 by formation of polymeric structures (Paradies, OligomericStructure of Mercury Cyanide in Solution, lecture before theGerman-Austrian Chemical Society, Innsbruck, May 1986). Thispharmaceutical preparation thus belongs to the substances of definedinterferon inductors of high molecular weight such as synthetic doublestranded RNA: PolyI:PolyC and of low molecular weight such as 10-carboxymethyl-9-acridanone. In spite of this heterogeneity of theinterferon action it is antivirally effective. This effect was used forthe biological detection of this pharmaceutical preparation. It can bestated that the interferon treatment of cells in the cell culture ismodified so that a subsequent virus replication in said cells isinhibited. Interferons thereby differ in their action mechanismfundamentally from virustatics which inhibit the metabolism of theviruses themselves. Since interferons act on the cells it is notsurprising that they can also exert other effects on the cells. Thisapplies not only to cell cultures but manifests itself in the entireorganism. It is known from a great variety of investigations thatinterferon inhibits the replication of a great number of viruses. Theextent of the inhibition depends on the particular virus system. Thus,the replication of almost all viruses appears to be subject toinhibition by interferon treatment of the cell. There are apparentlyvarious mechanisms by means of which interferons can become effective.Thus, for example, the replication of retroviruses is influenced by theformation of budding, i.e. the throwing out of newly formed viriones. Asimilar mechanism also appears to apply to the pharmaceuticalpreparation with micellarly included Hg(CN)₂. Thus, within the scope ofthe invention in the case of Herpes simplex virus (HSV 1-3 ) with thepharmaceutical preparation consisting of cetylpyridinium chloride andmercury cyanide (see example) the effect of induced interferon wasdetected on the synthesis of early viral-coded proteins of the HSv, theso-called β-proteins. In the case of the SV40 virus interferon alsoappears to act on a very early step of the replication lying even beforethe primary transcription.

The pharmaceutical preparation according to the invention, in particularmicellarly included mercury cyanide in7-hexadecylimidazolium-2,6-diamino-[4,5-d]-pyrimidine chloride,exhibited the interferon-induced inhibition in the case of various lyticRNA viruses. The inhibition takes place on the level of the regulationof the viral proteins. It is caused by induction of specific cellularenzymes. A more exact investigation showed that the enzymes inhibit the2',5'-oligoadenylate synthetase, the 2,5-A-dependent endoribonucleaseand the dsRNA-dependent protein kinase. By correlation studies andcharacterization of cell mutants it was possible to prove for the twoformer substances participation in the antiviral activity against lyticRNA viruses by micellarly bound Hg(CN)₂.

In these experimentally proved effects it was also possible to detect anantiproliferative effect of this pharmaceutical preparation on theinterferons in many ways on the membranes and on the cytoskeleton ofcells. Thus, they also influence the expressions of a number ofimportant genes, for example that of the main histocompatibility locus,the so-called transplantation antigens. Thus, immunological regulatoryeffects are also apparent (interleukin-1 synthesis). This gives thefollowing therapeutic and pharmacodynamic aspects: The induction of theinterferons by this pharmaceutical preparation leads to increasedexpression of the cell surface proteins which play the most importantpart in the immunological response. These are the so-calledtransplantation antigens. It should further be noted that at least twoimportant cellular components of the endogenous immune system areactivated, that is the macrophages and the natural killer cells. Also,in particular as regards interferon γ, the functions of the B-lymphocyteappear to be decisively influenced by this pharmaceutical preparation.

Thus, for the pharmaceutical preparation according to the invention, inparticular in the case of hexadecylpyridinium chloride and micellarlyincluded mercury cyanide or7-hexadecylimidazolium-2,6-diamino-[4,5-d]-pyrimidine chloride, or alsothe bromide, an induction of interferon results, although not a specificone. There can be no doubt that interferons have an immunologicalregulatory effect and not only an antiviral effect both in vitro and invivo. Although the direct antiviral effect can also be of significancein vivo, in the organism as regards the interferon effect as explainedabove other indirect mechanisms play a part, for example the activationof macrophages in particular in the case of the influenza virus A and B.The fact that interferon γ can activate macrophages also appears to beimportant with regard to bacterial and parasitical infections becausemacrophages play a significant part in the defence against suchinfections.

Posolgy and therapeutic indications

The therapeutic indications and the dose depend on the micellarlyincluded concentrations of the pharmaceutical active substance:

thus, indications exist for incipient and existing colds caused mainlybe influenza and rhino viruses;

catarrhal inflammations of viruidic origin;

skin infections and infectious dermatoses;

persistent antiseptic treatment of wounds;

dermatomycoses;

mycoses;

prophylaxis and therapy of bacterially induced skin lesions such aspyodermia, otitis media;

microbial and secondarily infected exzema;

oversensitivity to macrolide antibiotics;

acne, in particular inflammatory forms with papules and pustules;

bacterial infections and secondary infections of the skin in so far asthey are due to grampositive and/or gramnegative meclocycline-sensitivegerms;

acne vulgaris;

prevention of navel infections;

surgical and traumatic wounds;

local protection from infections and wounds infected withantibiotic-sensitive germs;

furuncles, carbuncles, abscesses;

dermatomycoses caused by dermatophytes, saccharomycetes, hyphomycetesand other fungi, pityriasis versicolor,

erythrasma through corne bacteria;

candidiaces of the skin and mucus membranes;

Herpes simplex I-III, Herpes Keratitis;

solar and senile keratoses, premalignant changes and superficialbasalioma, also in radiation-damaged skin;

squamous cell carcinoma of the skin and mucosa in the head and neckregion; dermatological malignant growths.

The specific dose depends on the diagnosis and pharmaceutical activesubstance. Since the maintenance and initial dose of the pharmaceuticalactive substances described here are known, depending on the type ofapplication and galenic preparation, e.g. creams, suppositories, drops,tablets, capsules and liposome-like encapsulations, only 50% of thenormal therapeutical total dose is required, depending on theconcentration of the micellarly included pharmaceutical activesubstance.

Outline of the dose reduction due to the potentiating (synergistic)effect

Micelles in aqueous solution, also those with included lipophilic activeagents, are in dynamic equilibrium with their monomeric tensides, i.e.the micelles change form, size and hydration. Under some circumstancesmonomeric cationic tensides leave a particular micelle to join againanother micelle in aqueous solution so that even when the concentrationof the tenside is far above the cmc there is always a certainconcentration of fluctuation monomers. By addition of the potentiatingmixture these dynamics are destroyed in that

(1) at constant temperature and chemical potential the form, size andmonodisperse homogeneity of the isotropic solution is retained andconsequently there is no loss of micellarly included lipophilic(hydrophobic) pharmaceutical active substance.

(2) The concentration of monomers which has a destablizing effect on thegeometrical form of the micelle is restricted in favour of incorporationinto the "complete" micelle in isotropic solution. As a result thesystem, including the micellarly included hydrophobic pharmaceuticalactive substances, "leaks". This is prevented mainly in that thepotentiating mixture, in particular glycerol and dimethylsulfoxidefreezes the water structure at the external surface of the micelle(tridymit structure) in such a manner that it assumes ice likestructures and the water molecules become very immobile.

(3) Due to the potentiating effect of the glycerol, as has beendemonstrated for example in vitro, the pharmaceutical preparation isless cytotoxic, i.e. it damages primarily the affected (infected) celland not so much the healthy cell in the cell unit.

The invention has in particular the following advantages:

The N-tensides prepared according to this invention, together with theinclusion according to the process of the invention of the activeagents, results in a considerable reduction, in some cases up to 80%, ofthe toxicity of the inorganic active agents or substances, e.g. withHg(CN)₂, (also HgCl₂, HG(NH₂)Cl [precipitate], ZnEDTA) and Zn salts ingeneral, as well as with nephrotoxic, ototoxic antibiotics, inparticular polymixins, erythromycin, gentamycin, tetracyclin, of about30% because

(1) the micelles, and their included active substances, are notresorbed, due to their size,

(2) the micellarly included active substances develop their effects onlyat the location, usually topically, so that small concentrations ofactive substances are adequate since additionally there is thesynergistic effect of the N-tenside.

It has thus been found inter alia that the inhibitory effect on salivarysecretion of atropine by hexadecylpyridinium chloride and bybenzothiazolium sulfate is intensified tenfold by micellar catalysiswith N-tensides made according to the process of the invention, pH≦7.0.The increased effect on the periphery is inter alia due to the micellarseparation of the racemate into L(-) hyocyamine (see FIG. 1).

Also, hexadecylbenzthiazolium sulfate for example stabilizes theatropine by incorporating the hydrophobic molecule regions of theatropine into the micellar core.

This content of the claim extends also to the antiphlogistic propertiesof the quaternary organic ammonium bases described here. The counter ionY⁻ has a process-inherent influence on the size, form and micellarstability but may also itself be a pharmaceutical active agent so that adrug action can be pharmacodynamically intensified. The tensidesdescribed here have the great advantage that in addition to theintrinsic pharmacodynamical properties they are environment-dependentand moreover stable in the acidic pH range. The micelles which can beobtained by the process as pharmaceutical preparation with includedlipophilic (hydrophobic) pharmaceutical active substances act ascarriers for antimicrobial, antiviral, keratolytic, antifungal andantineoplastic active substances but may also themselves inter alia havean antimicrobial, antifungal and antiviral and antiphlogistic (topical)effect.

In particular, the present invention describes a pharmaceuticalpreparation for providing micellar dissolved hydrophobic inorganicsubstances such as mercury II cyanide, zinc, tungsten and antimonycompounds as well as salts of phosphonic acid. It has been found thatthese organic compounds have both antiviral and antineoplastic effects.

The formation of the vesicular structures of the quaternary ammoniumbases of 4 and 3,5-substituted hexadecylpyridinium-Y⁻ also takes placespontaneously at constant temperature, pressure, ionic strength, also inthe presence of stoichiometrically pharmaceutical active substanceswhich can be bound both vesicularly (void) and micellarly (in the mannerof double membranes).

In particular the invention relates to an improved preparation methodfor making multilamella lipid bubbles on the basis of cationic tensideswhich can be used in particular to encapsulate lipophilic (hydrophobic)pharmaceutical active agents.

Most hitherto known processes suffer either from inadequateencapsulation effect or from a limitation of the types of materialswhich can be included or from both these defects. Thus, as is known mostof these processes are restricted to the inclusion of hydrophilicmaterials and pharmaceutical active substances and cannot efficientlyperform the inclusion of lipophilic pharmaceutical active substances. Incontrast thereto, all the presently available methods with the exceptionof that of Banghan et al. (Biochem. Biophys. Acta 443:629-634, 1976) aresuitable only for the encapsulation of biologically active substances inoligo-multilamella or unilamella liposomes.

A particular advantage of this pharmaceutical preparation on the basisof vesicular structures of N⁺ -tensides is the hydrophobic encapsulationof pharmaceutical active agents. A particularly advantageous consequenceof the large-size vesicles made by ultrasonic treatment and counter ionsis to be seen in that the danger of emergence of the pharmaceuticalactive substance from the bubble skin of the preparation is reduced oreliminated. Consequently, this form of the quaternary N⁺ -tensides onthe basis of six-member heterocycles can be employed in particular forencapsulation of hydrophobic pharmaceutical active substances which maybe used to achieve local, i.e. topically restricted, effects instead ofsystemic effects.

Whereas most of the known processes are restricted to the encapsulationof hydrophilic active agents with this invention encapsulation ofhydrophobic pharmaceutic active agents may be carried out. Tests haveshown that even inorganic lipophilic pharmaceutical active agents suchas mercury II cyanide can be included with high efficacy and theirpharmocodynamic effect can be further enhanced by the potentiatingmixture.

This disadvantage is eliminated by the novel N-tensides of the generalformula II and novel benzethonium compounds and also the vesicles on thebasis of N⁺ -tensides either by micellar inclusion of the pharmaceuticalactive substances or by covalent linking of the active substances to theN⁺ -tenside whilst retaining the outer morphological form of the overallmicelle.

The bactericidal effect of chlorhexidine on grampositive andgramnegative bacteria is known but gramnegative bacilli are resistant.It has been found that micellar solutions of quaternary ammonium basesaccording to the general formula I and in particular II which bond 2-4%by weight chlorhexidine hydrophobically in the micellar core cancel theresistance to gramnegative bacilli and increase their therapeuticalefficacy compared with chlorhexidine alone. The secondary effectsobserved of chlorhexidine such as contact dermatitis, skin effects oftopical nature, photosensibility of the skin, do not take place with themicellar solutions of the N-tensides of the formula I and II made by thepresent process.

It is the object of the present invention to eliminate the heterogeneitymentioned at the beginning of the form and size of the micelles even inthe presence of potentiating mixtures. It is thus ensured that amonodisperse form of cationic organic ammonium bases is achieved even inthe presence of pharmaceutical active substances and potentiatingmixtures in the preparation thereof.

These organic ammonium bases according to the invention obviate theaforementioned disadvantages of the hitherto known conventional invertsoaps. Thus, there is also great interest in the therapeutic use ofquaternary ammonium bases which function both as pharmaceutical activeagent and carrier of active agents of a great variety of types, forexample antimicrobial, antiviral, antifungal or antineoplastic nature,can absorb substances micellarly. They should therefore not have theaforementioned disadvantages dependent mainly on the environment.

The active substances covalently bound pharmaceutically, such aspyrimidine and purine derivatives at the N₁ or N₇, on the basis ofquaternary ammonium bases, have the advantage

1. that these masked antimetabolites from the pyrimidine or purineseries do not enter any intramolecular interactions of an anionic orcationic nature. They are neutrally charged (e.g. no nucleotide dianionby the phosphate) and can thus diffuse unrestrictedly into the pro oreukaryotic cell so that high intracellular antimetabolite (e.g.5'-nucleotide) concentrations are achieved;

2. that the pharmaceutical active substances by N-C-hydrolysis by meansof the enzyme systems present of the germinal or eukaryotic cells areliberated at the target or also topically;

3. by the increase in the hydrophobicity of the alkyl or aryl chain orthe radical at the N⁺ -tenside the membrane permeability is increased sothat the pharmaceutical active substances can pass quantitatively andpassively into the cytosol. In contrast to dianions or cations whichhave difficulty in passing through the membrane under physiological pHconditions and ionic strengths this can be done by the N⁺ -tensidesaccording to the invention without restriction;

4. the high hydrophobicity also gives a high distribution coefficient inthe system CHCl₃ -H₂ O at pH 8.0;

5. by the concentrated absorption of hydrophobic or hydrophilicpharmaceutical active substances, in addition to the covalently anchoredsubstances, the active substance concentration is increased afterpenetration through the germinal membrane, fungal cell wall (inhibitionof chitin synthetase) or viral phospholipid double membrane by aconcentration gradient (extracellular-intracellular). This results in alow flooding time.

In contrast to liposomes as carrier of pharmaceutical active substancesas described for example in U.S. Pat. No. 3,993,754 or European PatentNo. 0,102,324, the micelles of quaternary ammonium bases describedaccording to the invention have the advantages

1. that they can absorb water-insoluble active substances micellarly inthe so-called liquid core and as a result these water-insoluble activesubstances can be liberated in controlled manner both topically andenterally by opening of the micelle, said active substances being forexample rimantadine, amantadine, tromantadine, which are effectiveagainst influenza viruses and Herpes simplex viruses both of the skinand of the eye.

2. Water-soluble active substances can be dissolved both in the Sternlayer and also micellarly if they themselves have hydrophobic ranges,for example polyene compounds, tetracylines, aminoglycosides andaromatic antimetabolites, e.g. trifluorothymidine, viderabine,cytarabine, 5-iodo and 5-fluorodeoxyuridine, 5-ethyl-2'-deoxyuridine,erythromycin and nalidixic acid.

3. The cationic N-tensides according to the invention have two specificbonding sites with high bonding constants (K_(B) =10-15 μM) and highbonding capacity (capacity 100 μg/micelle): the first is due to thehydrophobic interaction between the liquid core of the micelle and thehydrophobic range of the active substance (ΔG=15-20 kcal/Mol) and alsothe π--π-interactions of the active substances described here betweenthe N-heterocycles of the N-tensides and the pharmaceutical activesubstances; the second binding site is nonspecific and is localized atthe interface between the Stern layer and the hydrophobic core. Thebinding constant lies in the region of K_(B) =20 mM and the bondingcapacity is 100-200 μg/micelle. The nonspecific binding sites are almostwithout exception in the Guy-Chapman layer. In contrast to liposomeswhere the number of nonspecific binding sites is substantially higherthe number of nonspecific binding sites can be eliminated by addition ofethanol/glycerol since the forces which act in the Guy-Chapman layer areeliminated by concentrations of ethanol and glycerol up to 30-35% byweight without influencing the bonding capacity and strength of thehydrophobic forces (only polarity and configuration).

4. The invention described here has the advantage that the micellarlyincluded or enclosed pharmaceutical active substances do not leave themicellar union as for example in the case of liposomes which with thehitherto known methods "leak". The sealing of the present invention ofmicellarly enclosed active substances can be detected for example inmicellarly bound radioactively marked trifluorothymidine, cytarabine andidoxuridine. It has been found inter alia that idoxuridine loses 5% byweight of its original micellarly included concentration (2000 μg) inthe case of hexadecylpyridinium or benzethonium chloride micelles onlyafter 200 days. The corresponding values for radioactively markedtrifluorothymidine and cytarabine are 210 and 300 days (20° C.).

5. According to the process of the present invention these micelles withthe included inorganic and organic active substances at pH=7.0 can bemade in simple manner without excessive apparatus expenditure in aqueousphase containing simple micelles with a diameter of about 50-100 Å andlarge micelles with a diameter of 600-10000 Å, depending on the counterion. In addition, by a mixture of glycerol/ethanol in a ratio of 2% byweight:15% by weight with respect to water both micelles of thedifferent order of magnitude are stabilized both in their form (sphere,hemicylinder, rod, disk) and in their compactness by lowering the cmc,as also by reduction of the free energy of the total micelle in theaqueous phase due to thinning of the electron density at the externalsurface. Small micelles can be preparatively separated from largemicelles by appropriate separation methods, e.g. HPLC, ultrafiltration,gel filtration and/or preparative centrifugation.

6. The stability, durability and storability of these micelles made inthis manner from quaternary organic ammonium bases, pH=7.0, totemperature, sealing and leaking and storage is increased by theincorporation of the pharmaceutical active substances into thehydrophobic core of the micelles compared with micelles without activesubstanccs for the same Y⁻. In contrast to the liposomes in this case nomelting occurs at higher temperature (>40° C.); with the preparationaccording to the invention the hydrodynamic conditions change onlyfrom >60° C. Since with increasing temperature the micelles ofquaternary ammonium bases made in this manner tend rather to undergo areduction in the hydrodynamic radius and therefore become more compact,this type of micelle is thermodynamically more stable than syntheticliposomes or liposomes+quaternary ammonium bases. These processes caneasily be checked by routine methods by inelastic light scattering intheir preparation.

7. The hydrophobicity or penetration of the H₂ O molecules into themicelles made in this manner and their influencing by inorganicpharmaceutical active substances, e.g. Hg(CN)₂, ZnEDTA, ZnSO₄, ZnO,wolframic acid antimonates, K₁₈ (KW₂₁ Sb₉ O₈₆)₁₇, and also organicsubstances, can be determined by NMR spectroscopy:

Taking as example 8-ketohexadecylpyridinium chloride (8-KHPCl) theincorporation of pharmaceutical active substances according to theinvention can be demonstrated. It has consequently now been found that achemical displacement of 146.6 ppm occurs for a 0.1 molar micellarsolution in water which is however shifted by for example Hg(CN)₂ to147.2 ppm. Micelles in aqueous solution which are 0.05 molar in 8-KHPCland 0.2 molar in CPCl (cetylpyridinium chloride) however gave a chemicaldisplacement of 147.2 ppm for the ¹³ C-carbonyl signal of 8-KHPCl. Ifthese two Figures are compared with the displacements of 8-KHPCI inmethanol (145.7 ppm) and acetonitrile (144.0 ppm) it becomes clear thatthe CO group in this micelle assumes a largely aqueous environment.Hg(CN)₂ plays here a double role which thus also governs the therapeuticwidth in vitro: the hydrophobic character of the Hg(CN)₂ effects a highsolubility in the hydrophobic core of for example hexadecylpyridiniumchloride as monomer and gives a chemical displacement of δ_(c8) 27.5 ppm¹³ C of CH₂ chain to 32.5 ppm whilst in 8-KHPCl micelles mercury IIcyanide is dissolved in the vicinity of the keto group (C₈) as Hg₂ (CN)₄in H₂ O (see above) and by this H₂ O solubility the concentration of H₂(CN)₄ is limited.

FIG. 3 shows the dependence of the extinction of the micellarly includedinorganic active substances and of the N-phosphono-acetyl-L-aspartate inhexadecylpyridinium chloride. Hereinafter a modification of theinvention is described which concerns in particular N-alkylatedquaternary nitrogen-containing heterocycles.

State of the art

Known are the quaternary ammonium bases with tenside-like effect of thegeneral structure (I) ##STR30## wherein generally

R₁ =an alkyl radical with 1-12 C atoms

R₂ =an alkyl radical with 1-12 C atoms

R_(n) =a straight-chain or branched alkyl radical with 10-20 C atoms oran alkeny radical with 8-10 C atoms or a 5 or 6-member aromaticheterocycle with one or 2 nitrogen atoms and

R_(m) =a straight-chain or branched alkyl radical with 10-20 C atoms oran alkenyl radical with 8-10 C atoms or a 5 or 6-member aromaticheterocycle with one or 2 nitrogen atoms

y⁻ =a monovalent anion

Compounds of this general formula have partly been described in theTensid-Taschenbuch, published by Dr. H. Stache, Carl-Hauser-Verlag,Munich, Vienna, 1981, pages 8/9.

Some of these compounds are also the object of a European patentapplication, application No. 83810338.0 of July 24, 1983, and thesetensides are used to prepare unilamellar liposomes in aqueous phase forthe dispersion.

It must also be taken into account here that these known N⁺ -tensides ofthe general formula I form both micellar and vesicular structures inaqueous and nonpolar solvents ((cf. for example J. Fendler, Acc. Chem.Res 1976, 9, 153; H. H. Paradies, J. Phys. Chem. 1986, 90, 5956; H. H.Paradies, 1982, Angew. Chem 10, 737; Angew. chem. Int. Ed. Engl. 1982,21, 765, Supplement 1982, 1670-1681) and here also micellarly catalyzedefined chemical and biophysical reactions depending on the objective.

In contrast, cationic tensides having a quaternary nitrogen within aπ-excess or π-defective aromatic, substituted or not substituted in thecore, are less well known. Descriptions exist for example forhexadecylpyridinium halide (cetylpyridinium halide), cf. inter aliaMerck Index 9, quinoline, cf. K. Lindner, inTenside-Textilhilfsstoffe-Waschrohstoffe 1964, volume 1, 987)andbenzthiazolium salts (European patent application No. 85400876.0 of May6, 1987, No. 660,802 Belgium, of Mar. 8, 1965) with various alkyl chainlengths and counterions for use in photography and electron transmissionby suitable formation of charge-transfer complexes. These are however2-methyl or 2-substituted benzthiazolium compounds with variablehydrophobic alkyl chain length of 12-30 carbon atoms at the heterocycleof the condensed-on benzene ring.

Furthermore, in the prior art the 2-substituted imidazolium salts and2-thiazolium compounds are described (cf. Tensid-Taschenbuch, H. Stache,2nd edition, 1981, pages 8/9), without however the cmc and othermicellar properties being specified. Corresponding matter is alsodescribed for the imidazolium compounds, cf. for exampleTensid-Textilhilfsmittel-Waschrohstoffe K. Lindner, 1964, 993;wissenschaftliche Verlagsgesellschaft, Stuttgart.

For vesicular compounds having a pyridine ring as aromatic substancesonly 4 and 3.5-alkyl or alkoxyl compounds have been described containinga methyl group at the quaternary nitrogen (cf. for example Sudholter etal. 1982, J. Amer. Chem. Soc. 104, 1069, Sudholter et al. 1979, J. Amer.Chem. Soc. 102, 2467).

The problem underlying the invention is to provide N-alkylatedquaternary nitrogen containing heterocycles.

This problem is solved by N-alkylated quaternary nitrogen-containingheterocycles, that is 2,5,6-substituted N₁ -alkylpyrimidinium compoundsof the formula ##STR31## wherein

n=8-20 in particular 15 and

Z⁻ =chloride, bromide, iodide, maleate, formate, acetate, propionate,hydrogen sulfate, malate, fumarate, salicylate, alginate, gluconate,glucoronate, galactoronate, ethyl sulfate or hydrogen phosphate H₂ PO₄⁻.

A preferred embodiment of the invention is the 2,5,6-substituted N₁-hexadecylpyrimidinium compounds of the formula ##STR32## wherein

Z⁻ =chloride, bromide, iodide, maleate, formate, acetate, propionate,hydrogen sulfate, malate, fumarate, salicylate, alginate, gluconate,glucoronate, galactoronate, ethyl sulfate or hydrogen phosphate H₂ PO₄⁻.

Preparation of the n-alkylated quaternary nitrogen containingheterocycles according to the invention (a). General remarks on thepreparation

These cationic tensides are characterized in that they have a very smallmicellization constant (cmc) of about 1.0-10⁻⁶ -1.5×10⁻⁷ mol/liter, havea very strong antimicrobial and antifungal effect, do not exhibitpolydispersity in the presence of inorganic anions or potentiatingmixtures and in some cases themselves are microbial metabolism products(antimetabolites) which are not toxic for the host cell.

The formation of the salt-like structure of this class of cationictensides of the form (HET.tbd.N⁺ -(CH₂)_(x) --CH₃) Y⁻ is due inter aliato the electron density distribution of the heteroaromatic nuclei and totheir basicity, including the influence of the substituents. A necessarycondition for the formation of quaternary salts of this five andsix-member heteroaromatic class is that the electron density at thenitrogen which is quaternized in accordance with MO-SCF calculationsmust have a magnitude of -0.08 (e.g. pyrazine-N₄) to -0.159 (e.g.imidazole-N₁, purine-N₇). This stability of the heterocyclic cationictensides described here depends also on its symmetry and chain length ofthe alkyl chain at the quaternary nitrogen.

In the case of imidazole, benzimidazole, for example stabilization iseffected by the formation of the salt and the quaternary nitrogen N₁ andthe free electron pair at the N₃ and the resulting high symmetry. Thesame applies to the H₉ -tautomers of purine and its symmetricallyarranged substituents which influence the negative charges at the N₁(-0.124) N₃ (-0.108) and N₉ (0.149) in such a manner that thequaternization at the N₉ is preferred in that the aforementioned orderN₁ →N₃ →N₉ is reversed. By the choice of suitable solvents the yieldscan be increased. Whereas for pyrdine, pyrimidine and imidazole radicalssymmetrical effects at the core play an important part, in the case forexample of pyrazine the electronic effect in the 2-position is ofsignificance but there are also very pronounced inductive effects (e.g.2-amino group), less than mesomers. This also applies to pyrazole.

The length of the alkyl chain at the quaternary nitrogen atom governsnot only the melting point and hydrophobicity of the cationic micellessubsequently formed in aqueous solution but in addition the yieldsdecrease with increasing chain length whilst the reaction times increasefor example in nitrobenzene or 2-ethoxyethanol.

Stable and easily crystallizable compounds are obtained for C₁₂ -C₁₈,the counter ion Y⁻ being always bromide and chloride. The othercompounds can easily be recrystallized from acetone or chloroform. Thecorresponding iodine compounds are temperature-sensitive andlight-sensitive.

(b) Specific remarks on the preparation

(1) Hexadecylpyrimidinium bromide, 0.01 mol 5-aminopyrimidine (0.95 g)and hexadecylbromide (0.01 mol (3.05 g) are reacted in 20 ml methanolwhilst stirring under nitrogen at 20° C. for 24 hours in the presence ofcatalytic amounts (0.5 mg) sodium amide. The resulting N₁-hexadecyl-5-aminopyrimidinium bromide is dissolved in acetone at 76° C.and after cooling to room temperature the N₁-hexadecyl-5-aminopyrimidinium bromide crystallizes out with a meltingpoint of 122° C. Yield 35%.

0.01 mol of this N₁ -hexadecyl-5-aminopyrimidinium bromide (3.20 g) isstirred for 6 hours under nitrogen in methanol/water ⁵⁰ /50 (^(v) /V) at0° C. in an ice bath with 1 g NaNO₂ and 0.1 ml concentrated hydrobromicacid. Thereafter the mixture is brought to room temperature and thenreflux heated at 80° C. for 2 hours under nitrogen whilst stirring. Theresulting hexadecylpyrimidinium bromide is extracted with2-ethoxyethanol and made to crystallize out at 10° C. Yield 30%, meltingpoint 105° C. (bromide), 189° C. (chloride).

Preparative separation of unreacted products can be achieved either byhigh-pressure liquid chromatography on an RP 18 column or by dialysisagainst phosphate-buffered aqueous solutions with subsequentDEAE-Sephadex A-50 exchanger chromatography of the residue from thedialysis tube.

In the UV spectrum of these N(1) alkylpyrimidinium salts, depending onthe substituent in 2,5 or 6-position, the maximum lies as a rule betweenλ_(max) =252 nm-265 nm.

In the ¹ H-NMR spectrum of the substituted or non-substituted N (1)alkylpyrimidinium halides prepared in CD Cl₃ /Me₄ Si apart from thecharacteristic signals of the corresponding pyrimidines there areadditionally observed the following characteristics: δ0.93 (3H, t, J=3.5Hz), 1.27 (nH, M); 5.05 (2H, m), 7.6-9.5 (3H, m, unsubstitutedpyrimidine).

In buffered aqueous solution below pH 7.0 (pH 3.9-5.9) for a micellesize of under 600 Å in diameter the typical sharp signals lie at δ about0.85-0.89 ppm (--CH₃), δ about 1.28-130 ppm (--CH₂ --) and about 3.25ppm (N--CH₂ --) for these N⁺ -tensides in addition to the protoncoupling signals of the unsubstituted pyrimidine core.

2. 2,5,6-position substituted pyrimidinium compounds are obtained byreaction in 2-ethoxyethanol under pressure in autoclaves at 100° C. fora reaction period of 8 hours from the corresponding n-alkyl bromides oriodides and the substituted pyrimidine compounds with yields between 30and 40%. The recrystallizations are carried out for all the substitutedpyrimidinium compounds from chloroform. The preparative separation ofunreacted products can be achieved as described above by high-pressureliquid chromatography. The N⁺ -tensides of the pyrimidines made in thismanner can also be obtained in uniform and pure form, free fromimpurities, by chromatographic separation on a DEAE-Sephadex A 25(1.5×30 cm) with a linear gradient of 0.1-0.5 MNaCl in 0.01 M₂ HPO₄, pH6.5.

3. N¹ -n-alkyl compounds of pyrimidine can be obtained by reaction ofn-alkyl-Mgx(x=Br, Cl) in good yields with pyrimidine or2,6,5,6-substituted pyrimidines in the presence of 1,2-dimethoxyethaneand/or n-heptane. No hetarin or addition-elimination orelimination-addition mechanism takes place.

0.01 mol (1.0 g) 5-fluoropyrimidine are dissolved in1,2-dimethoxymethane (100 ml) whilst stirring in a three-neck flaskunder nitrogen. From a dropping funnel 0.08 mol (same order of magnitudeas above) n-decylmagnesium chloride (or 0.09 mol≃29.6 gn-hexadecylmagnesium bromide), dissolved in 20 ml heptane, is addedslowly dropwise at 20° C. This solution is brought to 40° C., stirredfor 12 hours and when the reaction is completed from a dropping funnel20 ml of a 50% by weight solution of hydrobromic acid is added dropwiseat constant temperature. After 1 hour the excess Grignard is reacted. Itis cooled to 0° C. and any remaining excess of Grignard reagent isdestroyed by addition of methanol and the quaternary N₁ -pyrimidiniumbases extracted with 2-ethoxyethanol. The first recrystallization iscarried out from chloroform:methanol at 0° C. and the furtherrecrystallizations at room temperature.

Melting point: 5-fluoro-N₁ -decylpyrimidinium bromide 199° C. (dec.)

Melting point: 5-fluorohexadecylpyrimidinium bromide 175° C. (dec.)

Table 1 summarizes the characteristic properties of the N(1)-alkylatedquaternary pyrimidinium salts including their critical micellizationconcentration (cmc). Table 2 shows the hydrodynamic radii of these N⁺-tensides of the pyrimidines determined in dependence upon the counterions Z⁻.

                                      TABLE 1                                     __________________________________________________________________________    Characteristic properties of the N-alkylated quaternary pyrimidinium          compounds                                                                                                          Analysis (%) found                                                                        CMC × 10.sup.-6        NO.                                                                              N-Tenside              Z.sup.-                                                                            Fp °C.                                                                       C  H  N  Z  Mol/Liter                    __________________________________________________________________________    1. 2-Hydroxy-6-Amino-hexydecyl-pyrimidinium                                                             CL.sup.-                                                                           155   61,45                                                                            18,69                                                                            10,76                                                                            9,10                                                                             2,00                         2. Hexadecyl-pyrimidinium Br.sup.-                                                                           105   62,02                                                                            10,09                                                                            7,25                                                                             -- 2,50                         3. 2,6-Dihydroxy-5-Fluor-hexadecyl-pyrimidinium                                                         Cl.H.sub.2 O                                                                       172   61,13                                                                            22,68                                                                            7,14                                                                             9,05                                                                             0,85                         4. 2-Hydroxy-5-methyl-6-amino-hexydecyl-                                                                Br   192   56,18                                                                            26,67                                                                            9,36                                                                             -- 1,00                            pyrimidinium                                                               5. 2,6-Dihydro-dodecyl-pyrimidinium                                                                     Cl   195 (Zers)                                                                          64,79                                                                            13,78                                                                            9,45                                                                             -- 1,20                         __________________________________________________________________________

                                      TABLE 2                                     __________________________________________________________________________    Hydrodynamic radii of the N-tensides in dependence upon Z.sup.-               No.                                                                              N-Tenside              Z.sup.-                                                                             <R.sub.H >                                                                        (Å)                                   __________________________________________________________________________    1  2-Hydroxy-6-Amino hexadecyl pyrimidinium                                                             Cl.sup.-                                                                             50,0                                                                             +/-  10,0                                                           Br.sup.-                                                                            150,0                                                                             +/-  10,0                                                           H.sub.2 PO.sub.4 .sup.-                                                             150,0                                                                             +/-  10,0                                 2. Hexadecyl-pyrimidinium Cl.sup.-                                                                            450 +/-  20,0                                                           Br.sup.-                                                                            500 +/-  20,0                                                           Salicylate                                                                          1000                                                                              +/- 100,0                                 3. 2,6-Dihydroxy-5-Fluor-hexadecyl pyrimidinium                                                         Cl.sup.-                                                                            150,0                                                                             +/-  20,0                                                           Br.sup.-                                                                            200,0                                                                             +/-  20,0                                                           NO.sub.3 .sup.-                                                                      70,0                                                                             +/-  5,0                                                            H.sub.2 PO.sub.4 .sup.-                                                             1000,0                                                                            +/- 100,0                                 4. Dodecyl-pyrimidinium   Cl.sup.-                                                                            350,0                                                                             +/-  20,0                                                           Br.sup.-                                                                            480,0                                                                             +/-  20,0                                                           NO.sub.3 .sup.-                                                                     1000,0                                                                            + /-                                                                              100,0                                                           H.sub.2 PO.sub.4 .sup.-                                                             1000,0                                                                            +/- 150,0                                 5. N(1)-Octyl-2-Hydroxy-5-methyl-6-Amino-                                                               Br.sup.-                                                                            170,0                                                                             +/-  20                                      pyrimidinium           H.sub.2 PO.sub.4 .sup.-                                                             1000,0                                                                            +/- 150,0                                                           NO.sub.3 .sup.-                                                                     1500,0                                                                            +/- 200,0                                 __________________________________________________________________________

Uses:

The compounds newly described here surprisingly have a very small cmc inthe range of 10⁻⁵ -10⁻⁷ mol/liter which is largely independent of the pHand ionic strength (=0.1M). In addition, since in some cases they arethemselves antimetabolites, they have a biochemical and pharmacodynamiceffect. Due to their "monovalent" cationic nature they are able topenetrate the cell membranes of neoplastic tissues and then by unknownmechanisms after formation of the corresponding nucleosides andphosphorylation intervene in inhibitory manner in the transcription andin the translation. This is of particular significance for infectiousprocesses of bacterial (prophages) or above all viral nature.

Of significance for subsequent use of these N⁺ -tensides is the controlof the colloidal-chemical "aggregates" (micelles) by the counter ions Z⁻of both inorganic and organic nature. In contrast to many otheramphiphiles, which are not water-soluble, these N⁺ -tensides, due totheir hydrophobic effect, can form sheet-like double membrane structuresin water or aqueous solutions. Usually they have a cylindrical formwhich depends however very much on the counter ion: in the presence ofprimary phosphate (H₂ PO₄ ⁻) and in the presence of gluconate orgalactoronate highly oriented micellar fibril structures are formedwhich are stabilized by these anions. Thus, for example gel-likepreparations have helical structures on the basis of micellar cylinders.

I claim:
 1. 2,5,6-Substituted N₁ -hexadecylpyrimidinium compounds of theformula ##STR33## wherein Z.sup.⊖ =chloride, bromide, iodide, maleate,formate, acetate, propionate, hydrogen sulfate, malate fumarate,salicylate, alginate, gluconate, glucoronate, galactoronate, ethylsulfate or H₂ PO₄ ⁻.